JP2006523714A - Macrocyclic isoquinoline peptide inhibitor of hepatitis C virus - Google Patents

Abstract

［式中、R₁〜R₉、QおよびWは明細書に記載されている。］。該化合物を含む組成物および該化合物を用いてHCVを阻害する方法も開示する。Disclosed are compounds having formula 1, macrocyclic isoquinoline peptides:

[Wherein R _{1 to} R ₉ , Q and W are described in the specification. ]. Also disclosed are compositions comprising the compounds and methods of inhibiting HCV using the compounds.

Description

  The present invention generally comprises antiviral compounds, more specifically compounds that inhibit the function of NS3 protease (also referred to herein as “serine protease”) encoded by hepatitis C virus (HCV), Compositions comprising compounds and methods for inhibiting the function of NS3 protease are directed.

  HCV is the major human pathogen infecting an estimated 170 million people worldwide, about five times the number of human type 1 human immunodeficiency virus infections. A significant proportion of these HCV-infected individuals develop severe progressive liver disease, including cirrhosis and hepatocellular carcinoma (Lauer, GM; Walker, BDN Engl. J. Med. (2001), 345, 41-52 ).

  Currently, the most effective HCV therapy uses a combination of alpha-interferon and ribavirin and produces a lasting effect in 40% of patients (Poynard, T. et al. Lancet (1998), 352, 1426-1432). Recent clinical trial results demonstrate that polyethylene glycol pegylated alpha-interferon is superior to unmodified alpha-interferon monotherapy (Zeuzem, S. et al. N. Engl. J. Med) (2000), 343, 1666-1672). However, even with an experimental treatment regimen that includes a combination of polyethylene glycol-added alpha-interferon and ribavirin, a significant proportion of patients do not show a sustained reduction in viral load. That is, there is a clear and unmet medical need to develop effective therapies for the treatment of HCV infection.

  HCV is a positive strand RNA virus. Based on a comparison of the deduced amino acids and the extensive homology of the 5 'untranslated region, HCV has been classified into a distinct genus of the Flaviviridae family. All members of the Flaviviridae family have enveloped virions that contain positive-strand RNA genomes that encode all known virus-specific proteins via translation of a single continuous open reading frame.

  Considerable heterogeneity was found within the nucleotide and encoded amino acid sequence of the entire HCV genome. At least six major genotypes have been characterized and more than 50 subtypes have been described. The major genotypes of HCV vary globally, and the clinical significance of HCV genetic heterogeneity remains to be understood despite numerous studies on the etiology and possible effects of genotype on therapy. Hateful.

  The single-stranded HCV RNA genome is about 9500 nucleotides in length and has one open reading frame (ORF) that encodes one large polyprotein of about 3000 amino acids. In infected cells, this polyprotein is cleaved at multiple sites by cellular and viral proteases to produce structural and nonstructural (NS) proteins. In the case of HCV, the production of mature nonstructural proteins (NS2, NS3, NS4A, NS4B, NS5A, and NS5B) is performed by two viral proteases. The first one cleaves the NS2-NS3 junction, the second is a serine protease contained within the N-terminal region of NS3, cis at the NS3-NS4A cleavage site, and the remaining NS4A-NS4B, For NS4B-NS5A, NS5A-NS5B sites, trans (trans) mediates all subsequent cleavage downstream of NS3. NS4A protein appears to act as a cofactor for NS3 protease and may be involved in membrane localization of NS3 and other viral replicase components. Complex formation between the NS3 protease domain and its cofactor, NS4A, is essential for efficient proteolytic cleavage at each of the four sites. NS3 protein exhibits nucleoside triphosphatase and RNA helicase activity. NS5B is an RNA-dependent RNA polymerase that is involved in HCV replication.

Compounds that have been shown to have an inhibitory effect on HCV replication as selective HCV serine protease inhibitors (inhibitors) include macrocyclic peptide compounds disclosed in International Application PCT / CA00 / 00353 (Publication No. WO 00/59929).
(Summary of the Invention)

［式中、
(a) R₁、R₂、R₃、R₄、R₅、およびR₆は、それぞれ独立してH；C_1-6アルキル；C_3-7シクロアルキル；C_1-6アルコキシ；C_3-7シクロアルコキシ；ハロ-C_1-6アルコキシ；ハロ-C_1-6アルキル；シアノ；ハロ；ヒドロキシル；C_1-6アルカノイル；ニトロ；アミノ；モノもしくはジ-(C_1-6)アルキルアミン；モノもしくはジ-(C_3-7)シクロアルキルアミン；モノもしくはジ-C_1-6アルキルアミド；モノもしくはジ-(C_3-7)シクロアルキルアミド；カルボキシル；(C_1-6)カルボキエステル；チオール；C_1-6チオアルキル；C_1-6アルキルスルホキシド;C_1-6アルキルスルホン；C_1-6アルキルスルホンアミド；Hetで置換されていることがあるC_6-10アリール；C_7-14アルキルアリール；C_6-10アリールオキシ；C_7-14アルキルアリールオキシ；4-7員の単環式ヘテロアリールオキシ；またはHetである；該R₁〜R₆はC_1-6アルキル連結基によりイソキノリン基と結合していることがある;
(b) R₇はNH₂または-NR₁₀R₁₁である；ここで、R₁₀は、C_1-6アルキル、C_1-6ハロアルキル、C(O)-NR₁₂R₁₃、C(O)-OR₁₄、C(O)-SR₁₅、または-C(O)-R₁₆である；R₁₁は、H、C_1-6アルキル、またはC_1-6ハロアルキルである；ただし、R₁₂またはR₁₃のいずれかがHである場合はR₁₁はHである；
R₁₂およびR₁₃は、それぞれ独立してH；C_1-6アルキル、C_3-7シクロアルキル、またはC_4-10アルキルシクロアルキル(それぞれ、ハロ、C_1-3アルコキシ、C_1-3ハロアルコキシ、C_1-3アルキル、もしくはC_1-3ハロアルキルで置換されていることがある)；またはアリールである；ここで、R₁₂およびR₁₃は、それらが結合している窒素と一緒になって4-7員の複素環を形成し得る;
R₁₄およびR₁₅は、それぞれ独立してC_1-6アルキル、C_3-7シクロアルキル、またはC_4-10 アルキルシクロアルキル(それぞれハロ、C_1-3アルコキシ、C_1-3ハロアルコキシ、C_1-3アルキル、もしくはC_1-3ハロアルキルで置換されていることがある);アリール、またはHetである；R₁₆は、H；C_1-6アルキル、C_3-7シクロアルキル、またはC_4-10アルキルシクロアルキル(それぞれ、ハロ、C_1-3アルコキシ、C_1-3ハロアルコキシ、C_1-3アルキル、またはC_1-3ハロアルキルで置換されていることがある);アリールまたはHetである;
(c) R₈およびR₉は、それぞれ独立してH、またはハロで置換されていることがあるC_1-3アルキル、またはC_1-3アルコキシ、またはC_1-3ハロアルコキシである;
(d) Qは、O、S(O)_mから独立して選ばれる1〜3個の異種原子を含むことがある飽和または不飽和C_3-9鎖；(ここで、mは0、1、または2である)、またはNR₁₇(ここで、R₁₇はH；C_1-6アルキルまたはC_1-6シクロアルキル(それぞれがハロ、C_1-6アルコキシ、シアノ、またはC_1-6ハロアルコキシで置換されていることがある)；-C(O)-R₁₈、C(O)-OR₁₉、C(O)-NR₂₀R₂₁、または-SO₂R₂₂である；R₁₈、R₂₀、およびR₂₁は、それぞれ独立してH；C_1-6アルキル、またはC_1-6シクロアルキル(それぞれがハロ、C_1-6アルコキシ、シアノ、またはC_1-6ハロアルコキシで置換されていることがある)である；R₁₉は、C_1-6アルキルまたはC_1-6シクロアルキル(それぞれがハロ、C_1-6アルコキシ、シアノ、またはC_1-6ハロアルコキシで置換されていることがある)である;
R₂₂は、アリール、C_1-6アルキル、またはC_1-6シクロアルキル(それぞれがハロ、C_1-6アルコキシ、シアノ、またはC_1-6ハロアルコキシで置換されていることがある)である；
(e) Wは、OH、-NH-SO_n-R₂₃、またはNH-SO_n-R₂₄である；(ここでnは1または2である)、R₂₃は、C_1-8アルキル、C_4-10アルキルシクロアルキル、非置換C_3-7シクロアルキル、またはハロ、C_1-3アルコキシ、シアノ、アミン、モノもしくはジ-C_1-6アルキルアミン、モノもしくはジ-C_1-6アルキルアミド、またはカルボキシレートで置換されていることがあるC_7-9アルキルアリールまたはC_1-4アルキルで置換されていることがあるシクロプロピルもしくはシクロブチルである；R₂₄はC_6-10アリールまたはHetである。］。 The present invention provides a macrocyclic isoquinoline compound of the formula: or a pharmaceutically acceptable enantiomer, diastereomer, salt, solvate, or prodrug thereof.
Compounds of formula I:

[Where:
(a) R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , and R ₆ are each independently H; C _1-6 alkyl; C _3-7 cycloalkyl; C _1-6 alkoxy; C _{3 -7} cycloalkoxy; halo-C _1-6 alkoxy; halo-C _1-6 alkyl; cyano; halo; hydroxyl; C _1-6 alkanoyl; nitro; amino; mono- or di- (C _1-6 ) alkylamine; Mono or di- (C _3-7 ) cycloalkylamine; mono or di-C _1-6 alkylamide; mono or di- (C _3-7 ) cycloalkylamide; carboxyl; (C _1-6 ) carboxyester; thiol; C _1-6 thioalkyl; C _1-6 alkyl sulfoxide; C _1-6 alkyl sulfonate; C _1-6 alkyl sulfonamide; sometimes substituted with Het C _6-10 aryl; C _{7 - 14} alkyl Aryl; C _6-10 aryloxy; C _7-14 alkylaryloxy; 4-7 membered monocyclic heteroaryloxy; R _{1 to} R ₆ may be linked to an isoquinoline group via a C _1-6 alkyl linking group;
(b) R ₇ is NH ₂ or —NR ₁₀ R ₁₁ ; where R ₁₀ is C _1-6 alkyl, C _1-6 haloalkyl, C (O) —NR ₁₂ R ₁₃ , C (O) -OR ₁₄ , C (O) -SR ₁₅ , or -C (O) -R ₁₆ ; R ₁₁ is H, C _1-6 alkyl, or C _1-6 haloalkyl; provided that R ₁₂ or R ₁₁ is H when any of R ₁₃ is H;
R ₁₂ and R ₁₃ are each independently H; C _1-6 alkyl, C _3-7 cycloalkyl, or C _4-10 alkyl cycloalkyl (halo, C _1-3 alkoxy, C _1-3 halo, respectively) May be substituted with alkoxy, C _1-3 alkyl, or C _1-3 haloalkyl); or aryl; where R ₁₂ and R ₁₃ are taken together with the nitrogen to which they are attached. Can form a 4-7 membered heterocycle;
R ₁₄ and R ₁₅ are each independently C _1-6 alkyl, C _3-7 cycloalkyl, or C _4-10 alkyl cycloalkyl (halo, C _1-3 alkoxy, C _1-3 haloalkoxy, C May be substituted with _1-3 alkyl, or C _1-3 haloalkyl); aryl, or Het; R ₁₆ is H; C _1-6 alkyl, C _3-7 cycloalkyl, or C _{4 -10} alkylcycloalkyl (respectively, halo, C _1-3 alkoxy, sometimes C _1-3 haloalkoxy, substituted with C _1-3 alkyl or C _1-3 haloalkyl); is aryl or Het ;
(c) R ₈ and R ₉ are each independently C _1-3 alkyl, or C _1-3 alkoxy, or C _1-3 haloalkoxy, which may be substituted with H, or halo;
(d) Q is a saturated or unsaturated C _3-9 chain that may contain 1 to 3 heteroatoms independently selected from O, S (O) _m ; (where m is 0, 1 , Or 2), or NR ₁₇ where R ₁₇ is H; C _1-6 alkyl or C _1-6 cycloalkyl (each halo, C _1-6 alkoxy, cyano, or C _1-6 halo) May be substituted with alkoxy); —C (O) —R ₁₈ , C (O) —OR ₁₉ , C (O) —NR ₂₀ R ₂₁ , or —SO ₂ R ₂₂ ; R ₁₈ , R ₂₀ and R ₂₁ are each independently H; C _1-6 alkyl, or C _1-6 cycloalkyl (each substituted with halo, C _1-6 alkoxy, cyano, or C _1-6 haloalkoxy R ₁₉ is C _1-6 alkyl or C _1-6 cycloalkyl (each substituted with halo, C _1-6 alkoxy, cyano, or C _1-6 haloalkoxy) May be);
R ₂₂ is aryl, C _1-6 alkyl, or C _1-6 cycloalkyl, each optionally substituted with halo, C _1-6 alkoxy, cyano, or C _1-6 haloalkoxy. ;
(e) W is OH, —NH—SO _n —R ₂₃ , or NH—SO _n —R ₂₄ ; (where n is 1 or 2), R ₂₃ is C _1-8 alkyl, C _4-10 alkyl cycloalkyl, unsubstituted C _3-7 cycloalkyl, or halo, C _1-3 alkoxy, cyano, amine, mono or di-C _1-6 alkyl amine, mono or di-C _1-6 alkyl Amido, or C _7-9 alkylaryl optionally substituted with carboxylate or cyclopropyl or cyclobutyl optionally substituted with C _1-4 alkyl; R ₂₄ is C _6-10 aryl or Het It is. ].

  The invention also provides a composition comprising the compound or a pharmaceutically acceptable salt, solvate or prodrug thereof, and a pharmaceutically acceptable carrier. In particular, the present invention provides for inhibiting HCV NS3 protease comprising a therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt, solvate or prodrug thereof, and a pharmaceutically acceptable carrier. Useful pharmaceutical compositions are provided.

  The present invention further provides a method of treating an HCV-infected patient, comprising administering to the patient a therapeutically effective amount of a compound of the present invention, or a pharmaceutically acceptable salt, solvate, or prodrug thereof. Furthermore, this invention provides the method of inhibiting HCV NS3 protease by making HCV NS3 protease and a compound of this invention contact.

According to the present invention, it is now possible to provide an improved medicament containing the compound of the present invention that can be effective in the treatment of HCV-infected patients. Specifically, the present invention provides a peptide compound that can inhibit the function of the NS3 protease, for example, in combination with NS4A protease. In addition, the present invention makes it possible to administer combination therapy to patients, and the compounds of the present invention effective to inhibit HCV NS3 protease can be combined with other compounds having anti-HCV activity, such as HCV to treat HCV infection. To inhibit the function of a target selected from the group consisting of metalloprotease, HCV serine protease, HCV polymerase, HCV helicase, HCV NS4B protein, HCV entry, HCV assembly, HCV release, HCV NS5A protein, IMPDH and nucleoside analogues It can be administered with an effective compound.
(Detailed description of the invention)

  The stereochemical definitions and conventions used herein are generally described in the McGraw-Hill Dictionary of Chemical Terms, edited by SP Parker, McGraw-Hill Book Company, New York (1984), and Stereochemistry of Organic Compounds, Eliel, E. and Wilen. , S., John Wiley & Sons, Inc., New York (1994). Many organic compounds exist in optically active forms. That is, they have the ability to rotate plane polarized light. In the description of optically active compounds, the prefixes D and L or R and S are used to indicate the absolute configuration of the molecule about its chiral center. The prefix d and 1 or (+) and (-) means the compound is levorotatory (-) and the compound is dextrorotatory (+) or d Shows a representation of the rotation of plane polarized light by. For certain chemical structures, these compounds, called stereoisomers, are identical except that they are mirror images of one another. Certain stereoisomers of a mirror image pair are sometimes referred to as enantiomers, and mixtures of such isomers are often referred to as enantiomeric mixtures. When (R) or (S) is used, the absolute configuration of the substituents is indicated in the context of the entire compound rather than in the context of only the substituents.

  Unless otherwise stated herein, the following terms have the following definitions.

  The terms “racemic mixture” and “racemate” refer to an equimolar mixture of two enantiomeric species lacking optical activity.

  The term “chiral” refers to a molecule that has the property of non-superimposability of a mirror image partner, while the term “achiral” refers to a molecule that is superimposable with its mirror image partner.

  The term “stereoisomer” refers to compounds that have the same chemical composition but differ in the arrangement of atoms or groups in space.

  The term “diastereomers” refers to stereoisomers that are not enantiomers, for example, stereoisomers that have two or more centers of chirality and whose molecules are not mirror images of one another. Diastereomers have different physical properties, eg melting points, boiling points, spectral properties, and reactivity. Diastereomeric mixtures could be separated by high resolution quantitative methods such as electrophoresis and chromatography.

  The term “enantiomer” refers to two stereoisomers of a compound that are non-superimposable mirror images of each other.

  The term “therapeutically effective amount” means the total amount of each active compound sufficient to produce a meaningful patient benefit, eg, a sustained reduction in viral load. When applied to an individual active ingredient administered alone, the term refers only to that ingredient. When applied in combination, the term refers to the total amount of active ingredients that produce a therapeutic effect whether administered in combination, sequentially or simultaneously.

  The term “compound of the invention” and equivalent expressions are intended to include compounds of formula I, and pharmaceutically acceptable enantiomers, diastereomeric salts, and solvates, such as hydrates, and prodrugs thereof. To do. Similarly, for an intermediate, it is intended to include salts and solvates thereof where the context allows. Also included are preferred compounds with respect to the compounds of the invention, such as Formula II and A-M.

  The term “derivative” means a chemically modified compound, where modification is a routine procedure for chemists of ordinary skill, such as esters or amides of acids, protecting groups such as alcohols or It is considered that thiol is a benzyl group and amine is a tert-butoxycarbonyl group.

  The term “solvate” means a physical association of a compound of this invention with one or more organic or inorganic solvent molecules. This physical bond includes a hydrogen bond. In some cases, the solvate could be isolated, for example, when one or more solvent molecules are incorporated into the crystal lattice of the crystalline solid. “Solvate” encompasses both solution-phase and separable solvates. Examples of solvates include hydrates, ethanolates, methanolates, isopropanolates and the like.

  As used herein, the term “prodrug” refers to a compound of the invention that has a group that can be cleaved chemically or metabolically and that is pharmaceutically active in vivo by solvolysis or under physiological conditions. A derivative of the compound of the present invention. A prodrug of a compound could be formed by conventional methods using the functional group of the compound, eg, an amino, hydroxy, or carboxy group, if present. Prodrug derivative forms often provide solubility, histocompatibility, or delayed release advantages in mammalian organisms (see Bundgard, H., Design of Prodrugs, pp. 7-9, 21-24, Elsevier, Amsterdam 1985) . Prodrugs include acid derivatives well known to those skilled in the art, such as, for example, esters prepared by reaction of a parent acid compound with an appropriate alcohol, or amides prepared by reaction of a parent acid compound with an appropriate amine. Including.

  The term “patient” includes both humans and other mammals.

  The term “pharmaceutical composition” refers to the compound of the present invention at least one additional pharmaceutical carrier, ie adjuvant, excipient or vehicle, eg diluent, preservative, filler, flow rate, depending on the mode of administration and the nature of the dosage form. Means a composition comprising in combination with modifiers, disintegrants, humectants, emulsifiers, suspending agents, sweeteners, flavoring agents, fragrances, antibacterial agents, antiepileptic agents, lubricants, and dispensing agents. To do. For example, the ingredients described in Remington's Pharmaceutical Sciences, 18th Edition, Mack Publishing Company, Easton, PA (1999) could be used.

  As used herein, the term “pharmaceutically acceptable” refers to excessive toxicity, irritation, allergic reactions, or other problems or complications that are within reasonable medical judgment and balanced with a reasonable risk / benefit ratio. Compounds, substances, compositions suitable for use in contact with a patient's tissue for treatment without disease, such as the compounds and their enantiomers, diastereomers, salts, solvates, or prodrugs, and / or Or represents a dosage form.

  The term `` treating '' refers to (i) the occurrence of a disease, disorder, or condition in a patient who may not be diagnosed as yet, who may be susceptible to the disease, disorder, and / or condition. (Ii) inhibit the disease, disorder, or condition, i.e. stop its expression, (iii) reduce the disease, disorder, or condition, i.e., regress the disease, disorder, and / or condition Represents to bring.

  As used herein, the term “substituted” includes, unless otherwise specified, a substituent, eg, a mono-, di-, tri-, or tetra-substituted, core, such as an organic radical. From 1 to the maximum number of substitutions of the possible binding sites above are included.

  The nomenclature used to describe organic radicals such as hydrocarbons and substituted hydrocarbons generally follows standard nomenclature known in the art, unless otherwise specified. Group combinations, such as alkylalkoxyamines or arylalkyls, include all possible stable configurations unless otherwise specified. That is, the alkylaryl substituent can be attached to the core moiety by either the alkyl or aryl portion of the alkylaryl substituent, so long as the resulting compound is stable. Certain radicals and combinations are defined below for purposes of illustration.

  As used herein, the term “halo” refers to a halogen substituent selected from bromo, chloro, fluoro, or iodo. The term “haloalkyl” means an alkyl group substituted with one or more halo substituents.

As used herein, the term “alkyl” refers to an acyclic straight or branched chain hydrocarbon substituent of the specified number of carbons, such as methyl, ethyl, propyl, butyl, tert-butyl, hexyl, 1 -Methylethyl, 1-methylpropyl, 2-methylpropyl, 1,1-dimethylethyl are included. That is, C _1-6 alkyl represents an alkyl group having 1 to 6 carbon atoms. The term “lower alkyl” means an alkyl group having 1 to 6 carbon atoms, preferably 1 to 4 carbon atoms. The term “alkyl ester” refers to an alkyl group further comprising an ester group. In general, the ranges of carbon numbers indicated, for example C _2-6 alkyl esters, include all carbon atoms in the radical.

  As used herein, the term “alkenyl” refers to an alkyl radical containing at least one double bond, such as ethenyl (vinyl) and alkyl.

  As used herein, the term “alkoxy” refers to an alkyl group having the indicated number of carbon atoms attached to an oxygen atom. Alkoxy includes, for example, methoxy, ethoxy, propoxy, 1-methylethoxy, butoxy, and 1,1-dimethylethoxy. The latter radical is referred to in the art as tert-butoxy. The term “alkoxycarbonyl” means an alkoxy group further comprising a carbonyl group.

As used herein, the term “cycloalkyl” means a cyclic hydrocarbon substituent containing the indicated number of carbon atoms, eg, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and spirocyclic. Groups such as spirocyclopropyl and spirocyclobutyl are included. The term “cycloalkoxy” as used herein refers to a cycloalkyl group attached to an oxygen atom, for example, cyclobutyloxy or cyclopropyloxy. The term “alkylcycloalkyl” refers to a cycloalkyl group attached to an alkyl group. The ranges of carbon numbers shown include the total number of carbons in the radical unless otherwise specified. That is, a C _4-10 alkyl cycloalkyl could contain 1-7 carbon atoms in the alkyl group and 3-9 carbon atoms in the ring (eg, cyclopropylmethyl or cyclohexylethyl). .

As used herein, the term “aryl” means an aromatic moiety containing the indicated number of carbon atoms, such as, but not limited to, phenyl, indanyl, or naphthyl. For example, C _6-10 aryl represents an aromatic moiety having 6 to 10 carbon atoms that may be in the form of a monocyclic or bicyclic structure. The term “haloaryl” as used herein refers to an aryl that is mono-, di-, or tri-substituted with one or more halogen atoms. The terms “alkylaryl”, “arylalkyl”, and “aralkyl” mean an aryl group substituted with one or more alkyl groups. Unless the carbon range of each group is specified, the ranges described apply to total substitution. That is, the C _7-14 alkylaryl group has 1 to 8 carbon atoms in the alkyl group for monocyclic aromatics and 1 to 4 carbon atoms in the alkyl groups for fused aromatics. The bond of the group to the binding site on the molecule can be either an aryl group or an alkyl group. Unless a specific aryl radical is specified (eg, fluoro-phenyl), or unless the radical is described as unsubstituted, for example, an aryl radical in an aryl substituent or an alkylaryl substituent is known to those skilled in the art. typical substituents include halogen, hydroxy, carboxy, carbonyl, nitro, sulfo, amino, cyano, dialkylamino haloalkyl, CF _3, haloalkoxy, thioalkyl, alkanoyl, SH, alkylamino, alkylamido, dialkylamido, carboxyanhydride ester , Alkyl sulfones, alkyl sulfonamides, and those substituted with alkyl (alkoxy) amines. Examples of the alkylaryl group include benzyl, butylphenyl, and 1-naphthylmethyl.

As used herein, the term “alkanoyl” means a straight or branched 1-oxoalkyl radical containing the indicated number of carbon atoms, eg, formyl, acetyl, 1-oxopropyl (propionyl), 2 -Methyl-1-oxopropyl, 1-oxohexyl and the like.
As used herein, the term “alkylamide” refers to an amide monosubstituted with alkyl, such as

を意味する。

Means.

  As used herein, the term “heterocycle”, also referred to as “Het”, means a 7-12 membered bicyclic heterocycle and a 4-7 membered monocyclic heterocycle.

Preferred bicyclic heterocycles are 7-12 membered fused bicyclic systems containing 1 to 4 heteroatoms selected from nitrogen, oxygen, and sulfur (both rings share adjacent pairs of atoms). (Here, both rings of the heterocycle are completely unsaturated). Nitrogen and sulfur heteroatoms may be optionally oxidized. Bicyclic heterocycles may contain heteroatoms in one or both rings. Unless specified to a particular heterocycle (e.g., a 7-12 membered bicyclic heterocycle fluoride) or unless stated to be unsubstituted, the heterocycle includes the typical Those substituted with a substituent are included. For example, a bicyclic heterocycle may contain substituents, such as 1 to 3 substituents, on any ring carbon atom. Examples of suitable substituents include C _1-6 alkyl, C _3-7 cycloalkyl, C _1-6 alkoxy, C _3-7 cycloalkoxy, halo-C _1-6 alkyl, CF ₃ , mono- or di- -Halo-C _1-6 alkoxy, cyano, halo, thioalkyl, hydroxy, alkanoyl, NO ₂ , SH, amino, C _1-6 alkylamino, di (C _1-6 ) alkylamino, di (C _1-6 ) Alkyl amide, carboxyl, (C _1-6 ) carboxyester, C _1-6 alkyl sulfone, C _1-6 alkyl sulfonamide, C _1-6 alkyl sulfoxide, di (C _1-6 ) alkyl (alkoxy) amine, C Includes _6-10 aryl, C _7-14 alkylaryl, and 4-7 membered monocyclic heterocycles. When two substituents are attached to adjacent carbon atoms of a bicyclic heterocycle, they are joined to contain a ring, for example, up to two heteroatoms selected from oxygen and nitrogen 5, A 6- or 7-membered ring system can be formed. Bicyclic heterocycles are attached to the molecule at any atom in the ring, for example R _{1 of} formula I, preferably carbon.
Examples of bicyclic heterocycles include, but are not limited to, the following ring systems:

Preferred monocyclic heterocycles are 4-7 membered saturated, partially saturated, or fully unsaturated ring systems containing 1 to 4 heteroatoms selected from nitrogen, oxygen, and sulfur in the ring (this latter subset). Are also referred to herein as unsaturated heteroaromatics) (wherein the sulfur and nitrogen heteroatoms may be optionally oxidized). Unless a specific heterocycle is specified (e.g., C _1-6 alkoxy substituted 4-7 membered monocyclic heterocycle, or unless the heterocycle is described as unsaturated), the heterocycle is known to those skilled in the art. For example, a monocyclic heterocycle may contain substituents on any ring atom, such as 1 to 3 substituents. Examples of groups include C _1-6 alkyl, C _3-7 cycloalkyl, C _1-6 alkoxy, C _3-7 cycloalkoxy, halo-C _1-6 alkyl, CF ₃ , mono- or di-halo- C _1-6 alkoxy, cyano, halo, thioalkyl, hydroxy, alkanoyl, NO ₂ , SH, amino, C _1-6 alkylamino, di (C _1-6 ) alkylamino, di (C _1-6 ) alkylamide, carboxyl, (C _1-6) carboxyanhydride ester, C _1-6 alkyl sulfonate, C _1-6 alkyl sulfoxide, C _1-6 alkyl sulfonamidyl , Di (C _1-6) alkyl (alkoxy) amine, C _6-10 aryl, C _{7 - 14} alkylaryl, and a monocyclic heterocycle further 4-7 membered. Monocyclic heterocycle It may be attached to the molecule at any atom in the ring, for example R ₁ of formula I.

  Examples of monocyclic heterocycles include, but are not limited to, the following (and tautomers thereof):

  One skilled in the art will recognize that the heterocycle used in the compounds of the present invention should be stable. In general, stable compounds are those that can be synthesized, isolated, and formulated using techniques known to those skilled in the art without degradation of the compound.

  The term “substituent” with respect to an amino acid or amino acid derivative means a radical derived from the corresponding α-amino acid. For example, the substituents methyl, iso-propyl, and phenyl represent the amino acids alanine, valine, and phenylglycine, respectively.

  The designations “P1 ′, P1, P2, P3, and P4” used herein for naming compounds of the present invention are the amino acid residues of the protease inhibitor that bind to the binding of the natural peptide cleavage substrate. Map the relative position of. Cleavage occurs on the natural substrate between P1 and P1 ′, where the non-prime position displays the amino acid starting from the C-terminus of the peptide's natural cleavage site and extending toward the N-terminus, while the prime location is the cleavage site. It originates from the indicated N-terminus and extends towards the C-terminus. For example, P1 ′ represents the first position outside the right-hand end of the C-terminal of the cleavage site (i.e., the N-terminal first position), but P1 starts numbering from the left-hand side of the C-terminal cleavage site (P2 : 2nd position from C-terminal etc.) [See Berger A. & Schechter I., Transactions of the Royal Society London series (1970), B257, 249 264].

  According to the present invention, a compound represented by the following formula, or a pharmaceutically acceptable enantiomer, diastereomer, salt, solvate, or prodrug thereof is provided.

［式中、
(a) R₁、R₂、R₃、R₄、R₅、およびR₆は、それぞれ独立してH；C_1-6アルキル；C_3-7シクロアルキル；C_1-6アルコキシ；C_3-7シクロアルコキシ；ハロ-C_1-6アルコキシ；ハロ-C_1-6アルキル；シアノ；ハロ；ヒドロキシル；C_1-6アルカノイル；ニトロ；アミノ；モノもしくはジ-(C_1-6)アルキルアミン；モノもしくはジ-(C_3-7)シクロアルキルアミン；モノもしくはジ-C_1-6アルキルアミド；モノもしくはジ-(C_3-7)シクロアルキルアミド；カルボキシル；(C_1-6)カルボキエステル；チオール；C_1-6チオアルキル；C_1-6アルキルスルホキシド;C_1-6アルキルスルホン；C_1-6アルキルスルホンアミド；Hetで置換されていることがあるC_6-10アリール；C_7-14アルキルアリール；C_6-10アリールオキシ；C_7-14アルキルアリールオキシ；4-7員の単環式ヘテロアリールオキシ；またはHetである；該R₁〜R₆はC_1-6アルキル連結基によりイソキノリン基と結合していることがある;
(b) R₇はNH₂または-NR₁₀R₁₁である；ここで、R₁₀は、C_1-6アルキル、C_1-6ハロアルキル、C(O)-NR₁₂R₁₃、C(O)-OR₁₄、C(O)-SR₁₅、または-C(O)-R₁₆である；R₁₁は、H、C_1-6アルキル、またはC_1-6ハロアルキルである；ただし、R₁₂またはR₁₃のいずれかがHである場合はR₁₁はHである；
R₁₂およびR₁₃は、それぞれ独立してH；C_1-6アルキル、C_3-7シクロアルキル、またはC_4-10アルキルシクロアルキル(それぞれ、ハロ、C_1-3アルコキシ、C_1-3ハロアルコキシ、C_1-3アルキル、もしくはC_1-3ハロアルキルで置換されていることがある)；またはアリールである；ここで、R₁₂およびR₁₃は、それらが結合している窒素と一緒になって4-7員の複素環を形成し得る;
R₁₄およびR₁₅は、それぞれ独立してC_1-6アルキル、C_3-7シクロアルキル、またはC_4-10 アルキルシクロアルキル(それぞれハロ、C_1-3アルコキシ、C_1-3ハロアルコキシ、C_1-3アルキル、もしくはC_1-3ハロアルキルで置換されていることがある);アリール、またはHetである；R₁₆は、H；C_1-6アルキル、C_3-7シクロアルキル、またはC_4-10アルキルシクロアルキル(それぞれ、ハロ、C_1-3アルコキシ、C_1-3ハロアルコキシ、C_1-3アルキル、またはC_1-3ハロアルキルで置換されていることがある);アリールまたはHetである;
(c) R₈およびR₉は、それぞれ独立してH、またはハロで置換されていることがあるC_1-3アルキル、またはC_1-3アルコキシ、またはC_1-3ハロアルコキシである;
(d) Qは、O、S(O)_mから独立して選ばれる1〜3個の異種原子を含むことがある飽和または不飽和C_3-9鎖；(ここで、mは0、1、または2である)、またはNR₁₇(ここで、R₁₇はH；C_1-6アルキルまたはC_1-6シクロアルキル(それぞれがハロ、C_1-6アルコキシ、シアノ、またはC_1-6ハロアルコキシで置換されていることがある)；-C(O)-R₁₈、C(O)-OR₁₉、C(O)-NR₂₀R₂₁、または-SO₂R₂₂である；R₁₈、R₂₀、およびR₂₁は、それぞれ独立してH；C_1-6アルキル、またはC_1-6シクロアルキル(それぞれがハロ、C_1-6アルコキシ、シアノ、またはC_1-6ハロアルコキシで置換されていることがある)である；R₁₉は、C_1-6アルキルまたはC_1-6シクロアルキル(それぞれがハロ、C_1-6アルコキシ、シアノ、またはC_1-6ハロアルコキシで置換されていることがある)である;
R₂₂は、アリール、C_1-6アルキル、またはC_1-6シクロアルキル(それぞれがハロ、C_1-6アルコキシ、シアノ、またはC_1-6ハロアルコキシで置換されていることがある)である；
(e) Wは、OH、-NH-SO_n-R₂₃、またはNH-SO_n-R₂₄である；(ここでnは1または2である)、R₂₃は、C_1-8アルキル、C_4-10アルキルシクロアルキル、非置換C_3-7シクロアルキル、またはハロ、C_1-3アルコキシ、シアノ、アミン、モノもしくはジ-C_1-6アルキルアミン、モノもしくはジ-C_1-6アルキルアミド、またはカルボキシレートで置換されていることがあるC_7-9アルキルアリールまたはC_1-4アルキルで置換されていることがあるシクロプロピルもしくはシクロブチルである；R₂₄はC_6-10アリールまたはHetである。］。
好ましくは、R₁がC₃位に結合し、H；C_1-6アルキル；C_3-7シクロアルキル；C_1-6アルコキシ；C_3-7シクロアルコキシ；ハロ-C_1-6アルコキシ；ハロ-C_1-6アルキル；シアノ；ハロ；C_1-6アルカノイル；モノもしくはジ-(C_1-6)アルキルアミン；モノもしくはジ-C_1-6アルキルアミド；カルボキシル；Hetで置換されていることがあるC_6-10アリール；C_7-14アルキルアリール；C_6-10アリールオキシまたはHetから選ばれる。好ましくは、R₂、R₃、およびR₄がそれぞれC₄、C₅、およびC₆位に結合し、それぞれ独立してH；C_1-6アルキル；C_3-7シクロアルキル；C_1-6アルコキシ；C_3-7シクロアルコキシ；ハロ-C_1-6アルコキシ；ハロ-C_1-6アルキル；シアノ；ハロ；ヒドロキシ；C_1-6アルカノイル；モノもしくはジ-(C_1-6)アルキルアミン；モノもしくはジ-(C_3-7)シクロアルキルアミン；モノもしくはジ-C_1-6アルキルアミド；モノもしくはジ-(C_3-7)シクロアルキルアミド；カルボキシル；Hetで置換されていることがあるC_6-10アリール；C_7-14アルキルアリール；C_6-10アリールオキシ；またはHetから選ばれる。好ましくは、R₅およびR₆がそれぞれC₇およびC₈位に結合し、それぞれ独立してH；C_1-3アルキル；C_3-4シクロアルキル；C_1-3アルコキシ；C_3-4シクロアルコキシ；ハロ-C_1-3アルコキシ；ハロ-C_1-3アルキル；シアノ；ハロ；ヒドロキシ；C_1-3アルカノイル；モノもしくはジ-(C_1-3)アルキルアミン；モノもしくはジ-(C_3-4)シクロアルキルアミン；モノもしくはジ-C_1-3アルキルアミド；モノもしくはジ-(C_3-4)シクロアルキルアミド；またはカルボキシルから選ばれる。所望により、R₁、R₂、R₃、R₄、R₅、およびR₆置換基の1またはそれ以上がC_1-6アルキル連結基、例えばメチレン基によりイソキノリン基と結合することができる。
好ましくは、Qが、O、S(O)_mから独立して選ばれる1〜3個の異種原子を含むことがある飽和または不飽和C_3-9鎖；(ここでmは0、1、または2である)、またはNR₁₇であり、R₁₇がH；C_1-6アルキル、C_1-6シクロアルキル、-C(O)-R₁₈、C(O)-OR₁₉、C(O)-NR₂₀R₂₁、または-SO₂R₂₂である。好ましくは、R₁₈、R₂₀、およびR₂₁が、それぞれ独立してH；C_1-6アルキル、またはC_1-6シクロアルキルであり；R₁₉がC_1-6アルキルまたはC_1-6シクロアルキルであり、R₂₂がアリール、C_1-6アルキル、またはC_1-6シクロアルキル(それぞれがハロで置換されていることがある)である。 Formula I:

[Where:
(a) R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , and R ₆ are each independently H; C _1-6 alkyl; C _3-7 cycloalkyl; C _1-6 alkoxy; C _{3 -7} cycloalkoxy; halo-C _1-6 alkoxy; halo-C _1-6 alkyl; cyano; halo; hydroxyl; C _1-6 alkanoyl; nitro; amino; mono- or di- (C _1-6 ) alkylamine; Mono or di- (C _3-7 ) cycloalkylamine; mono or di-C _1-6 alkylamide; mono or di- (C _3-7 ) cycloalkylamide; carboxyl; (C _1-6 ) carboxyester; thiol; C _1-6 thioalkyl; C _1-6 alkyl sulfoxide; C _1-6 alkyl sulfonate; C _1-6 alkyl sulfonamide; sometimes substituted with Het C _6-10 aryl; C _{7 - 14} alkyl Aryl; C _6-10 aryloxy; C _7-14 alkylaryloxy; 4-7 membered monocyclic heteroaryloxy; R _{1 to} R ₆ may be linked to an isoquinoline group via a C _1-6 alkyl linking group;
(b) R ₇ is NH ₂ or —NR ₁₀ R ₁₁ ; where R ₁₀ is C _1-6 alkyl, C _1-6 haloalkyl, C (O) —NR ₁₂ R ₁₃ , C (O) -OR ₁₄ , C (O) -SR ₁₅ , or -C (O) -R ₁₆ ; R ₁₁ is H, C _1-6 alkyl, or C _1-6 haloalkyl; provided that R ₁₂ or R ₁₁ is H when any of R ₁₃ is H;
R ₁₂ and R ₁₃ are each independently H; C _1-6 alkyl, C _3-7 cycloalkyl, or C _4-10 alkyl cycloalkyl (halo, C _1-3 alkoxy, C _1-3 halo, respectively) May be substituted with alkoxy, C _1-3 alkyl, or C _1-3 haloalkyl); or aryl; where R ₁₂ and R ₁₃ are taken together with the nitrogen to which they are attached. Can form a 4-7 membered heterocycle;
R ₁₄ and R ₁₅ are each independently C _1-6 alkyl, C _3-7 cycloalkyl, or C _4-10 alkyl cycloalkyl (halo, C _1-3 alkoxy, C _1-3 haloalkoxy, C May be substituted with _1-3 alkyl, or C _1-3 haloalkyl); aryl, or Het; R ₁₆ is H; C _1-6 alkyl, C _3-7 cycloalkyl, or C _{4 -10} alkylcycloalkyl (respectively, halo, C _1-3 alkoxy, sometimes C _1-3 haloalkoxy, substituted with C _1-3 alkyl or C _1-3 haloalkyl); is aryl or Het ;
(c) R ₈ and R ₉ are each independently C _1-3 alkyl, or C _1-3 alkoxy, or C _1-3 haloalkoxy, which may be substituted with H, or halo;
(d) Q is a saturated or unsaturated C _3-9 chain that may contain 1 to 3 heteroatoms independently selected from O, S (O) _m ; (where m is 0, 1 , Or 2), or NR ₁₇ where R ₁₇ is H; C _1-6 alkyl or C _1-6 cycloalkyl (each halo, C _1-6 alkoxy, cyano, or C _1-6 halo) May be substituted with alkoxy); —C (O) —R ₁₈ , C (O) —OR ₁₉ , C (O) —NR ₂₀ R ₂₁ , or —SO ₂ R ₂₂ ; R ₁₈ , R ₂₀ and R ₂₁ are each independently H; C _1-6 alkyl, or C _1-6 cycloalkyl (each substituted with halo, C _1-6 alkoxy, cyano, or C _1-6 haloalkoxy R ₁₉ is C _1-6 alkyl or C _1-6 cycloalkyl (each substituted with halo, C _1-6 alkoxy, cyano, or C _1-6 haloalkoxy) May be);
R ₂₂ is aryl, C _1-6 alkyl, or C _1-6 cycloalkyl, each optionally substituted with halo, C _1-6 alkoxy, cyano, or C _1-6 haloalkoxy. ;
(e) W is OH, —NH—SO _n —R ₂₃ , or NH—SO _n —R ₂₄ ; (where n is 1 or 2), R ₂₃ is C _1-8 alkyl, C _4-10 alkyl cycloalkyl, unsubstituted C _3-7 cycloalkyl, or halo, C _1-3 alkoxy, cyano, amine, mono or di-C _1-6 alkyl amine, mono or di-C _1-6 alkyl Amido, or C _7-9 alkylaryl optionally substituted with carboxylate or cyclopropyl or cyclobutyl optionally substituted with C _1-4 alkyl; R ₂₄ is C _6-10 aryl or Het It is. ].
Preferably, R ₁ is bonded to the C ₃ position, H; C _1-6 alkyl; C _3-7 cycloalkyl; C _1-6 alkoxy; C _3-7 cycloalkoxy; halo-C _1-6 alkoxy; -C _1-6 alkyl; cyano; halo; C _1-6 alkanoyl; mono- or di- (C _1-6 ) alkylamine; mono- or di-C _1-6 alkylamide; carboxyl; selected from C _6-10 aryloxy or Het; is C _6-10 aryl; C _{7 - 14} alkylaryl. Preferably, R ₂ , R ₃ , and R ₄ are bonded to the C ₄ , C ₅ , and C ₆ positions, respectively, and are independently H; C _1-6 alkyl; C _3-7 cycloalkyl; C _{1- 6} alkoxy; C _3-7 cycloalkoxy; halo-C _1-6 alkoxy; halo-C _1-6 alkyl; cyano; halo; hydroxy; C _1-6 alkanoyl; mono- or di- (C _1-6 ) alkylamine Mono or di- (C _3-7 ) cycloalkylamine; mono or di-C _1-6 alkylamide; mono or di- (C _3-7 ) cycloalkylamide; carboxyl; may be substituted with Het; Selected from C _6-10 aryl; C _7-14 alkyl aryl; C _6-10 aryloxy; or Het. Preferably, R ₅ and R ₆ are bonded to the C ₇ and C ₈ positions, respectively, and are independently H; C _1-3 alkyl; C _3-4 cycloalkyl; C _1-3 alkoxy; C _3-4 cyclo alkoxy; halo -C _1-3 alkoxy; halo -C _1-3 alkyl; cyano; halo; hydroxy; C _1-3 alkanoyl; mono- or di - (C _1-3) alkyl amine; mono- or di - (C _{3 -4)} cycloalkyl amine; mono- or di -C _1-3 alkyl amide; mono- or di - (C _3-4) cycloalkyl amide; selected from or carboxyl. If desired, one or more of the R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , and R ₆ substituents can be attached to the isoquinoline group by a C _1-6 alkyl linking group, such as a methylene group.
Preferably, Q is a saturated or unsaturated C _3-9 chain that may contain 1 to 3 heteroatoms independently selected from O, S (O) _m ; (where m is 0, 1, Or NR ₁₇ and R ₁₇ is H; C _1-6 alkyl, C _1-6 cycloalkyl, —C (O) —R ₁₈ , C (O) —OR ₁₉ , C (O ) -NR ₂₀ R ₂₁ , or -SO ₂ R ₂₂ . Preferably, R ₁₈ , R ₂₀ , and R ₂₁ are each independently H; C _1-6 alkyl or C _1-6 cycloalkyl; R ₁₉ is C _1-6 alkyl or C _1-6 cyclo Alkyl and R ₂₂ is aryl, C _1-6 alkyl, or C _1-6 cycloalkyl, each of which may be substituted with halo.

Preferably, W is OH, —NH—SO _n —R ₂₃ , or NH—SO _n —R ₂₄ , n is 1 or 2, and R ₂₃ is unsubstituted C _3-7 cycloalkyl, or C ₇ Cyclopropyl or cyclobutyl which may be substituted with _-9 alkylaryl or C _1-4 alkyl, and R ₂₄ is C _6-10 aryl or Het.

で示される化合物、またはその医薬的に許容されるエナンチオマー、ジアステレオマー、塩、溶媒和物、またはプロドラッグを提供する
［式中、
(a) R₁は、H；C_1-6アルキル；C_3-7シクロアルキル；C_1-6アルコキシ；C_3-7シクロアルコキシ；ハロ-C_1-6アルコキシ；ハロ-C_1-6アルキル；シアノ；ハロ；C_1-6アルカノイル；モノもしくはジ-(C_1-6)アルキルアミン；モノもしくはジ-C_1-6アルキルアミド；カルボキシル；Hetで置換されていることがあるC_6-10アリール；C_7-14アルキルアリール；C_6-10アリールオキシまたはHetである；該R₁はC_1-6アルキル連結基によりイソキノリン基と結合していることがある；
R₂、R₃、およびR₄は、それぞれ独立してH；C_1-6アルキル；C_3-7シクロアルキル；C_1-6アルコキシ；C_3-7シクロアルコキシ；ハロ-C_1-6アルコキシ；ハロ-C_1-6アルキル；シアノ；ハロ；ヒドロキシル；C_1-6アルカノイル；モノもしくはジ-(C_1-6)アルキルアミン；モノもしくはジ-(C_3-7)シクロアルキルアミン；モノもしくはジ-C_1-6アルキルアミド；モノもしくはジ-(C_3-7)シクロアルキルアミド；カルボキシル；Hetで置換されていることがあるC_6-10アリール；C_7-14アルキルアリール；C_6-10アリールオキシ；またはHetである；該R₂〜R₄はC_1-3アルキル連結基によりイソキノリン基と結合していることがある；R₅およびR₆は、それぞれ独立してH；C_1-3アルキル；C_3-4シクロアルキル；C_1-3アルコキシ；C_3-4シクロアルコキシ；ハロ-C_1-3アルコキシ；ハロ-C_1-3アルキル；シアノ；ハロ；ヒドロキシル；C_1-3アルカノイル；モノもしくはジ-(C_1-3)アルキルアミン；モノもしくはジ-(C_3-4)シクロアルキルアミン；モノもしくはジ-C_1-3アルキルアミド；モノもしくはジ-(C_3-4)シクロアルキルアミド；またはカルボキシルである;
(b) R₇はNH₂または-NR₁₀R₁₁である；R₁₀はC_1-6アルキル、C_1-6ハロアルキル、C(O)-NR₁₂R₁₃、C(O)-OR₁₄、または-C(O)-R₁₆である；R₁₁はH、C_1-6アルキル、またはC_1-6ハロアルキルである；ただし、R₁₂またはR₁₃のいずれかがHである場合はR₁₁はHである；R₁₂およびR₁₃は、それぞれ独立してH；C_1-6アルキル、C_3-7シクロアルキル、またはC_4-10アルキルシクロアルキル(それぞれがハロ、C_1-3アルコキシ、C_1-3ハロアルコキシ、C_1-3アルキル、またはC_1-3ハロアルキルで置換されていることがある)であり；R₁₂およびR₁₃は、それらが結合している窒素と一緒になって4-7員の複素環を形成し得る；R₁₄およびR₁₅は、それぞれ独立してC_1-6アルキル、C_3-7シクロアルキル、またはC_4-10アルキルシクロアルキル(それぞれがハロ、C_1-3アルコキシ、C_1-3ハロアルコキシ、C_1-3アルキル、またはC_1-3ハロアルキルで置換されていることがある)である；R₁₆は、H；C_1-6アルキル、C_3-7シクロアルキル、またはC_4-10アルキルシクロアルキル(それぞれがハロ、C_1-3アルコキシ、C_1-3ハロアルコキシ、C_1-3アルキル、またはC_1-3ハロアルキルで置換されていることがある);アリールまたはHetである;
(c) R₈およびR₉は、それぞれ独立してH、またはハロで置換されていることがあるC_1-3アルキル、またはC_1-3アルコキシ、またはC_1-3ハロアルコキシである;
(d) Qは、O、S(O)_mから独立して選ばれる1〜3個の異種原子を含むことがある飽和または不飽和C_3-9鎖である；(ここで、mは0、1、または2である)、またはNR₁₇である、ここで、R₁₇は、H；C_1-6アルキル、C_1-6シクロアルキル、-C(O)-R₁₈、C(O)-OR₁₉、C(O)-NR₂₀R₂₁、または-SO₂R₂₂である；R₁₈、R₂₀、およびR₂₁は、それぞれ独立してH；C_1-6アルキルまたはC_1-6シクロアルキルである；R₁₉はC_1-6アルキルまたはC_1-6シクロアルキルである；R₂₂はアリール、C_1-6アルキル、またはC_1-6シクロアルキル(それぞれがハロで置換されていることがある)である；
(e) Wは、OH、-NH-SO_n-R₂₃、またはNH-SO_n-R₂₄である、ここで、nは1または2である；R₂₃は、非置換C_3-7シクロアルキル、またはC_7-9アルキルアリールもしくはC_1-4アルキルで置換されていることがあるシクロプロピルまたはシクロブチルである；R₂₄はC_6-10アリールまたはHetである。］。 According to one aspect of the present invention, Formula II:

Or a pharmaceutically acceptable enantiomer, diastereomer, salt, solvate, or prodrug thereof [wherein
(a) R ₁ is H; C _1-6 alkyl; C _3-7 cycloalkyl; C _1-6 alkoxy; C _3-7 cycloalkoxy; halo-C _1-6 alkoxy; halo-C _1-6 alkyl cyano, halo, C _1-6 alkanoyl; mono- or di - (C _1-6) alkyl amine; C _6-10 that may have been substituted with Het; mono- or di -C _1-6 alkyl amide; carboxyl Aryl; C _7-14 alkylaryl; C _6-10 aryloxy or Het; the R ₁ may be linked to the isoquinoline group by a C _1-6 alkyl linking group;
R ₂ , R ₃ , and R ₄ are each independently H; C _1-6 alkyl; C _3-7 cycloalkyl; C _1-6 alkoxy; C _3-7 cycloalkoxy; halo-C _1-6 alkoxy Halo-C _1-6 alkyl; cyano; halo; hydroxyl; C _1-6 alkanoyl; mono- or di- (C _1-6 ) alkylamine; mono- or di- (C _3-7 ) cycloalkylamine; Di-C _1-6 alkylamide; mono- or di- (C _3-7 ) cycloalkylamide; carboxyl; C _6-10 aryl optionally substituted with Het; C _7-14 alkylaryl; C _{6- 10} aryloxy; or Het; the R _{2 to} R ₄ may be linked to the isoquinoline group by a C _1-3 alkyl linking group; R ₅ and R ₆ are each independently H; C _{1 -3} alkyl; C _3-4 cycloalkyl; C _1-3 alkoxy; C _3-4 cycloalkoxy; halo-C _1-3 alkoxy; halo-C _1-3 a Cyano; halo; hydroxyl; C _1-3 alkanoyl; mono- or di- (C _1-3 ) alkylamine; mono- or di- (C _3-4 ) cycloalkylamine; mono- or di-C _1-3 alkyl Amide; mono- or di- (C _3-4 ) cycloalkylamide; or carboxyl;
(b) R ₇ is NH ₂ or —NR ₁₀ R ₁₁ ; R ₁₀ is C _1-6 alkyl, C _1-6 haloalkyl, C (O) —NR ₁₂ R ₁₃ , C (O) —OR ₁₄ , or is _{-C (O) -R 16; R} 11 is H, is C _1-6 alkyl or C _1-6 haloalkyl; provided that when either R ₁₂ or R ₁₃ is H R ₁₁ R ₁₂ and R ₁₃ are each independently H; C _1-6 alkyl, C _3-7 cycloalkyl, or C _4-10 alkyl cycloalkyl (each halo, C _1-3 alkoxy, May be substituted with C _1-3 haloalkoxy, C _1-3 alkyl, or C _1-3 haloalkyl); R ₁₂ and R ₁₃ together with the nitrogen to which they are attached 4-14 membered heterocycles may be formed; R ₁₄ and R ₁₅ are each independently C _1-6 alkyl, C _3-7 cycloalkyl, or C _4-10 alkyl cycloalkyl (each halo, C _1-3 alkoxy, C _1-3 haloalkoxy, C _1-3 Alkyl or it is there) substituted with C _1-3 haloalkyl,; R ₁₆ is, H; C _1-6 alkyl, C _3-7 cycloalkyl or C _4-10 alkylcycloalkyl (each, May be substituted with halo, C _1-3 alkoxy, C _1-3 haloalkoxy, C _1-3 alkyl, or C _1-3 haloalkyl); aryl or Het;
(c) R ₈ and R ₉ are each independently C _1-3 alkyl, or C _1-3 alkoxy, or C _1-3 haloalkoxy, which may be substituted with H, or halo;
(d) Q is a saturated or unsaturated C _3-9 chain that may contain 1 to 3 heteroatoms independently selected from O, S (O) _m ; (where m is 0 , 1 or 2), or NR ₁₇ where R ₁₇ is H; C _1-6 alkyl, C _1-6 cycloalkyl, —C (O) —R ₁₈ , C (O) —OR ₁₉ , C (O) —NR ₂₀ R ₂₁ , or —SO ₂ R ₂₂ ; R ₁₈ , R ₂₀ , and R ₂₁ are each independently H; C _1-6 alkyl or C _1-6 R ₁₉ is C _1-6 alkyl or C _1-6 cycloalkyl; R ₂₂ is aryl, C _1-6 alkyl, or C _1-6 cycloalkyl, each substituted with halo May be);
(e) W is OH, —NH—SO _n —R ₂₃ , or NH—SO _n —R ₂₄ , where n is 1 or 2; R ₂₃ is unsubstituted C _3-7 cyclo Alkyl, or cyclopropyl or cyclobutyl optionally substituted with C _7-9 alkylaryl or C _1-4 alkyl; R ₂₄ is C _6-10 aryl or Het. ].

Preferably, R ₁ is H; C _1-3 alkoxy; mono- or di- (C _1-6 ) alkylamine; 5- or 6-membered monocyclic heterocycle; or 5- or 6-membered monocyclic heterocycle C _6-10 aryl which may be substituted with Preferably, R ₂ , R ₃ , R ₄ , and R ₅ are each independently H; C _1-6 alkoxy; halo-C _1-6 alkoxy; hydroxyl; or mono- or di- (C _1-6 ) Alkylamine.

Preferably, R ₇ is NH ₂ or —NHR ₁₀ ; R ₁₀ is C (O) —NR ₁₂ R ₁₃ or C (O) —OR ₁₄ ; R ₁₂ and R ₁₃ are substituted with halo Sometimes C _1-6 alkyl; R ₁₄ is C _1-6 alkyl or C _3-7 cycloalkyl, optionally substituted with halo.

［式中、Pは、O、S(O)_m(ここで、mは0、1、または2である)から独立して選ばれる1個の異種原子を含むことがある飽和C₃鎖、またはNR₁₇(式IIの化合物の場合と同意義である)ある。]。 Preferably, C is a C _5-7 membered chain having one double bond that may contain one heteroatom independently selected from O, S (O) _m , where m is 0 , 1, or 2), or NR ₁₇ (where R ₁₇ is H; C _1-6 alkyl or C _1-6 cycloalkyl). In a preferred aspect of the invention, Q has the following structure:

Wherein P is a saturated C ₃ chain that may contain one hetero atom independently selected from O, S (O) _m, where m is 0, 1, or 2. Or NR ₁₇ (same as in the case of the compound of formula II). ].

In certain aspects of the invention, W is —NH—SO _n —R ₂₃ (where n is 1 or 2), and R ₂₃ is unsubstituted C _3-7 cycloalkyl, or C _7-9 alkyl Preference is given to cyclopropyl or cyclobutyl which may be substituted with aryl or C _1-4 alkyl. In another aspect of the invention, W is NH—SO _n —R ₂₄ , n is 1 or 2, and R ₂₄ is Het.

からなる群から選ばれる。 Preferably, when R ₂₄ is Het, Het is

Selected from the group consisting of

で示される化合物、またはその医薬的に許容されるエナンチオマー、ジアステレオマー、塩、溶媒和物、またはプロドラッグを提供する
［式中、
(a) R₁は、H；C_1-3アルコキシ；ジ-(C_1-6)アルキルアミン；5または6員の単環式複素環；または5または6員の単環式複素環で置換されていることがあるC_6-10アリールである；R₂、R₃、R₄、およびR₅は、それぞれ独立してH；C_1-3アルコキシ；ハロ；またはジ-(C_1-6)アルキルアミンである;
(b) R₇は-NHR₁₀である；ここで、R₁₀はC(O)-NHR₁₃またはC(O)-OR₁₄である；R₁₃およびR₁₄はC_1-6アルキルである；
(c) Qは、独立してO、S(O)_mから選ばれる1個の異種原子を含むことがある1個の二重結合を有するC_5-7員の鎖(ここで、mは0、1、または2である)、またはNR₁₇(ここで、R₁₇はH；C_1-6アルキルまたはC_1-6シクロアルキルである)である；
(d) R₂₃は、非置換C_3-7シクロアルキル、またはC_7-9アルキルアリールまたはC_1-4アルキルで置換されていることがあるシクロプロピルまたはシクロブチルである。］。
好ましくは、R₁が、ピリジン、ピロリジン、モルホリン、ピペラジン、オキサゾール、イソキサゾール、チアゾール、イミダゾール、ピロール、およびピラゾールからなる群から選ばれる。ある好ましい局面において、R₁が、C_1-3アルコキシ、ハロ、カルボキシル、ジ-(C_1-3)アルキルアミン、C_1-3ハロアルキル、トリフルオロメチル、トリフルオロメトキシ、およびヒドロキシからなる群から選ばれる1またはそれ以上のメンバーで置換されていることがあるフェニルである。別の好ましい局面において、R₁がジ-(C_1-3)アルキルアミンである。別の好ましい局面において、R₁が、C_1-3アルキル、C_5-7シクロアルキル、またはピリジンからなる群から選ばれる1またはそれ以上のメンバーで置換されたピペラジンである。好ましくは、R₂がクロロまたはフルオロである。ある好ましい局面において、R₂がジ-(C_1-3)アルキルアミンまたはメトキシである。 In another aspect of the invention, Formula III:

Or a pharmaceutically acceptable enantiomer, diastereomer, salt, solvate, or prodrug thereof [wherein
(a) R ₁ is H; C _1-3 alkoxy; di- (C _1-6 ) alkylamine; 5- or 6-membered monocyclic heterocycle; or 5- or 6-membered monocyclic heterocycle is is C _6-10 aryl that may occur _{are; R 2, R 3, R} 4, and R ₅ are each independently H; C _1-3 alkoxy; halo; or di - (C _1-6 Alkylamine;
(b) R ₇ is —NHR ₁₀ ; where R ₁₀ is C (O) —NHR ₁₃ or C (O) —OR ₁₄ ; R ₁₃ and R ₁₄ are C _1-6 alkyl;
(c) Q is a C _5-7 membered chain having one double bond that may contain one heteroatom independently selected from O and S (O) _m (where m is Is 0, 1, or 2), or NR ₁₇ where R ₁₇ is H; C _1-6 alkyl or C _1-6 cycloalkyl;
(d) R ₂₃ is unsubstituted C _3-7 cycloalkyl, or cyclopropyl or cyclobutyl which may be substituted with C _7-9 alkylaryl or C _1-4 alkyl. ].
Preferably, R ₁ is selected from the group consisting of pyridine, pyrrolidine, morpholine, piperazine, oxazole, isoxazole, thiazole, imidazole, pyrrole, and pyrazole. In certain preferred aspects, R ₁ is from the group consisting of C _1-3 alkoxy, halo, carboxyl, di- (C _1-3 ) alkylamine, C _1-3 haloalkyl, trifluoromethyl, trifluoromethoxy, and hydroxy. A phenyl that may be substituted with one or more members of choice. In another preferred aspect, R _{1 is} di- (C _1-3 ) alkylamine. In another preferred aspect, R ₁ is piperazine substituted with one or more members selected from the group consisting of C _1-3 alkyl, C _5-7 cycloalkyl, or pyridine. Preferably R ₂ is chloro or fluoro. In certain preferred aspects, R _{2 is} di- (C _1-3 ) alkylamine or methoxy.

から選ばれる構造を有する。 In certain preferred aspects, Q is

Having a structure selected from

から選ばれる構造を有する。 In another preferred aspect, Q is

Having a structure selected from

からなる群から選ばれる。 Examples of preferred compounds of the present invention are:

Selected from the group consisting of

  The compound of the present invention containing a basic moiety can form a salt by adding a pharmaceutically acceptable acid. Acid addition salts are compounds of Formula I, and pharmaceutically acceptable inorganic or organic acids including, but not limited to, hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid, For example, it is formed from p-toluenesulfonic acid, methanesulfonic acid, acetic acid, benzoic acid, citric acid, malonic acid, fumaric acid, oxalic acid, succinic acid, sulfamic acid, or tartaric acid. That is, examples of such pharmaceutically acceptable salts include chloride, bromide, iodide, sulfate, phosphate, methanesulfonate, citrate, acetate, malonate, fumarate, Sulfamate and tartrate are included.

  Amine group salts may include quaternary ammonium salts in which the amino nitrogen carries a suitable organic group, such as an alkyl, alkenyl, alkynyl, or aralkyl moiety.

  Compounds of the invention that are substituted with an acidic group could exist as salts formed through base addition. Such base addition salts include those derived from inorganic bases, such as alkali metal salts (eg, sodium and potassium), alkaline earth metal salts (eg, calcium and magnesium), aluminum salts and ammonium. Contains salt. In addition, suitable base addition salts include pharmaceutically acceptable organic salts such as trimethylamine, triethylamine, morpholine, pyridine, piperidine, picoline, bicyclohexylamine, N, N′-dibenzylethylenediamine, 2-hydroxyethylamine, Bis (2-hydroxyethyl) amine, tri (2-hydroxyethyl) amine, procaine, dibenzylpiperidine, N-benzyl-β-phenethylamine, dehydroabiethylamine, N, N'-bishydroabiethylamine, glucamine, N- Methylglucamine, collidine, quinine, quinoline, ethylenediamine, ornithine, choline, N, N'-benzylphenethylamine, chloroprocaine, diethanolamine, diethylamine, piperazine, tris (hydroxymethyl) aminomethane, and tetramethylammonium Dorokishido, as well as basic amino acids, such as lysine, arginine, and salts of N- methyl-glutamine. These salts may be prepared by methods known to those skilled in the art.

  A list of suitable salts can be found, for example, in Remington's Pharmaceutical Sciences, 18th Edition, Mack Publishing Company, Easton, PA, 1990, p. 1445.

  Certain compounds of the invention and their salts exist in the form of solvates with water, such as hydrates, or with organic solvents such as methanol, ethanol, or acetonitrile, respectively methanolate, ethanolate, or acetonitrile. A rate could be formed. The present invention includes each solvate and mixtures thereof.

  Furthermore, the compounds of the invention or salts or solvates thereof may exhibit polymorphism. The present invention also includes all such polymorphic forms.

  The compounds of the invention (formula I, II or III) also contain two or more chiral centers and exist in different optically active forms. For example, a compound of formula I could contain the cyclopropyl group shown in the P1 moiety below.

［式中、C₁およびC₂は、それぞれシクロプロピル環の1および2位の不斉炭素原子を示す。本発明化合物の他の部分に他の考えられる不斉中心に関わらず、これら2個の不斉中心の存在は、該化合物がQが下記のようにアミドに対してsynまたはカルボニルに対してsynに配置する式IIIの化合物のジアステレオマーのようなジアステレオマーの混合物として存在しうることを意味する。

[Wherein, C ₁ and C ₂ represent asymmetric carbon atoms at positions 1 and 2 of the cyclopropyl ring, respectively. Regardless of other possible asymmetric centers in other parts of the compounds of the invention, the presence of these two asymmetric centers means that the compound is synthesized to Q for amide or carbonyl for carbonyl as described below. Means that it may exist as a mixture of diastereomers, such as diastereomers of the compound of formula III located in

  The present invention includes both enantiomers and enantiomeric mixtures of such compounds, for example racemic mixtures.

  Enantiomers are resolved by methods known to those skilled in the art, for example by formation of diastereomeric isomer salts and then separated by crystallization, gas-liquid or liquid chromatography, selective reaction of one enantiomer with an enantiomer specific reagent. I can do it. It will be appreciated that when the desired enantiomer is converted to another chemical component by separation techniques, additional steps are required to form the desired enantiomeric form. Alternatively, specific enantiomers could be synthesized by asymmetric synthesis using optically active reagents, substrates, catalysts, or solvents, or by converting one enantiomer to another by asymmetric transformation.

  Certain compounds of the present invention may also exist in different stable steric forms that are separable. For example, torsional asymmetry due to rotational restrictions on asymmetric single bonds due to steric hindrance or ring strain may allow separation of different conformers. The present invention includes each conformational isomer of these compounds and mixtures thereof.

  Certain compounds of the present invention may exist in zwitterionic forms and the present invention includes zwitterionic forms of these compounds and mixtures thereof.

  The starting materials useful for the synthesis of the compounds of the invention are known to those skilled in the art and can be readily prepared or are commercially available.

  The compounds of the present invention can be prepared by methods known to those skilled in the art. The following methods are illustrative and do not limit the scope of the present invention. For example, a compound of the present invention having a structure of formula I, II, or III can be synthesized from a compound of formula IV, formula VIIA, or VIIB as shown in the following reaction scheme. It will be appreciated that it may be preferred or necessary to prepare the compound in which the functional group is protected with a conventional protecting group and then the protecting group is removed to yield a compound of the invention. Details regarding the use of protecting groups in the present invention are known to those skilled in the art.

The α-carboxylic acid of the compound of formula VIIA or VIIB is in the presence of a peptide coupling agent such as CDI or EDAC, and a base such as 4-dimethylaminopyridine (4-DMAP) and / or 1,8-diazabicyclo [5.4. .0] Coupling with R ₂₃ SO ₂ NH ₂ (R ₂₃ is as defined above) in the presence of undeca-7-ene (DBU) to form a compound of formula I, II, or III. In the construction of compounds of formula I, the P1 ′ end is incorporated into the molecule as detailed below using one of the general methods described above. In certain instances, the P1 ′ element, which is a cycloalkylsulfonamide, alkylsulfonamide, or heteroarylsulfonamide, is commercially available or the corresponding alkyl- or cycloalkyl-sulfonyl chloride by treating the sulfonyl chloride with ammonia. Can be manufactured from. Alternatively, these sulfonamides can be synthesized using a general method described in the following reaction formula. In it, the commercially available 3-chloro-propylsulfonyl chloride (1) is converted into an appropriately protected sulfonamide, for example by treatment with tert-butylamine. The resulting sulfonamide (2) is then converted to the corresponding cycloalkylsulfonamide by treatment with two equivalents of a base such as butyllithium in a solvent such as THF at low temperature. The resulting cycloalkylsulfonamide can be deprotected by treatment with an acid to obtain the desired unprotected cycloalkylsulfonamide. The P1 ′ moiety can be incorporated into a compound of formula I.

  Compounds having the structure of formula IV, VI, VIIA, or VIIB are described, for example, in this specification and in International Application No. PCT / US01 / 45145, Publication No. WO 02/060926 (published August 8, 2002); Application No. PCT / CA00 / 00353 (publication No. WO 00/59929, publication date 12 October 2000); and US Pat. No. 6,323,180 (patent November 27, 2001).

  Compounds having the structure of Formula IV can also be prepared by coupling chemical precursor A with chemical precursor B as follows.

  The chemical precursor B can be synthesized, for example, as follows.

  Treatment of a commercially available or easily synthesized imine (B1) with 1,4-dihalobutene (B2) in the presence of a base gives the imine (B3). Next, B having a syn vinyl substituent with respect to the carboxyl group is obtained by acid hydrolysis of B3. For compounds B3 and B, the vinyl group is preferably syn to the ester. The amine moiety of B can be protected with a Boc group to give the fully protected amino acid B4. This intermediate is a racemate that can be resolved by an enzymatic step that cleaves the ester portion of B4 to give the corresponding carboxylic acid. Without being bound by any particular theory, the reaction is such that one of the enantiomers B4 (1S, 2R) undergoes a reaction at a much higher rate than its mirror image B4 (1R, 2S), and the kinetic resolution of the intermediate racemate. To be selective. In the above example, the more preferred stereoisomer for integrating into the tripeptide is B4 (1R, 2S). In the presence of the enzyme, this enantiomer does not undergo ester cleavage and this enantiomer B5 is recovered from the reaction mixture. However, the less preferred enantiomer B4 (1S, 2R) undergoes ester cleavage, ie enzyme-catalyzed hydrolysis, to give the free acid B6. The ester (B5) can be separated from the acid product (B6) by, for example, an aqueous extraction method or a lutinic method such as chromatography.

  Compounds of formula 1 can also be converted to other compounds of formula I as described herein. An example of such a method is the following reaction formula, where a compound of formula I (1) holding a Boc group at the P4 position is converted to a compound of formula I (3) holding a urea group at the P4 position: Is done. Conversion of (1) to (3) can be performed in a two-step process, initially by treating (1) with an acid such as TFA in a solvent such as methylene chloride (1). ) To amine (2). The resulting amine TFA salt can be treated with an isocyanate in the presence of 1 equivalent of a base to give a compound of formula I (3) in which the P3 moiety is covered with urea. As indicated above, one skilled in the art will recognize that intermediate (2) can be used as a starting material for preparing compounds of formula I in which the P3 group is covered with an amide or carbamate. The construction of the compound of formula I can be achieved using standard conditions to form the P4 functionality from an amine.

  The procedure for preparing the P2 intermediate and the compound of formula I is shown in the following reaction scheme. It should be noted that in many cases the reaction is only shown for one position of the intermediate. However, it should be understood that the reaction may be used to modify other locations within the intermediate. Furthermore, the intermediates, reaction conditions, and methods shown in the specific examples are widely applicable to compounds with other substitution patterns. The following general reaction scheme is followed by the examples herein. The general and specific examples are not limiting.

In the above reaction scheme, an N-protected C4-hydroxyproline moiety and an isoquinoline heterocycle are coupled to form an intermediate (4), and then the peptide extension method described herein and the ring-closing olefin metathesis reaction. Shown is modification of intermediate (4) to a compound of formula I. It should be noted that a base is used in the first step of coupling the C4-hydroxyproline group with the isoquinoline element. One skilled in the art will recognize that this coupling can be performed with a base such as potassium tert-butoxide or sodium hydride in a solvent such as DMF or DMSO or THF. This coupling with the isoquinoline ring system occurs at the C1 position (numbering for the isoquinoline ring system shown in Intermediate 2) and is indicated by a chloro group that replaces this method. It should be noted that other leaving groups can be utilized at this position, for example fluoro, as shown in the reaction scheme. The fluoro intermediate (3) is available from the corresponding chloro compound using literature methods described herein.
In some of the examples herein, isoquinoline is incorporated into the final compound, specifically the P2 region of the compound. One skilled in the art will appreciate that many general methods are available for the synthesis of isoquinolines. Furthermore, the isoquinoline produced by these methods can be easily incorporated into the final compound of formula I using the methods described herein. One general methodology for synthesizing isoquinoline is shown in the following reaction scheme. Here, the cinnamic acid derivative represented by structure (2) in general form is converted to 1-chloroisoquinoline in a four-step process.

次いで、該クロロイソキノリンを本明細書に記載のごとくカップリング反応に用いC4-ヒドロキシプロリン誘導体を得ることができる。桂皮酸のクロロキノリンの変換は塩基の存在下で桂皮酸をアルキルクロロホルメートで処理して開始する。次いで、得られた無水物をアジ化ナトリウムで処理して該本発明に示すアシルアジド(3)の形成を得る。代わりの方法をカルボン酸からアシルアジドの形成に利用可能であり、例えば該カルボン酸を塩基存在下で塩化メチレンのような非プロトン性溶媒中のジフェニルホスホリルアジド(DPPA)で処理することができる。反応手順の次の工程において、アシルアジド(3)を該反応式に示すように対応するイソキノロン(4)に変換する。その場合、アシルアジドをジフェニルメタンのような高沸点溶媒中で約190℃の温度に加熱する。この反応は一般的で、対応する桂皮酸誘導体から中程度〜高収率で置換イソキノロンをもたらす。該桂皮酸誘導体は市販されているか、マロン酸またはその誘導体と直接縮合し、またWittig反応を用いることにより対応するベンズアルデヒド(1)誘導体から得ることができることに留意すべきである。中間体イソキノロン(4)をオキシ塩化リンで処理し、対応する1-クロロイソキノリンに変換することができる。この反応は一般的であり、これを本明細書に記載のあらゆるイソキノロンに適用してヒドロキシ置換基を対応するクロロ化合物に変換することができる。

The chloroisoquinoline can then be used in a coupling reaction as described herein to obtain a C4-hydroxyproline derivative. The conversion of cinnamic acid to chloroquinoline begins by treating cinnamic acid with alkyl chloroformate in the presence of a base. The resulting anhydride is then treated with sodium azide to give the acyl azide (3) formation shown in the present invention. Alternative methods are available for the formation of acyl azides from carboxylic acids, for example the carboxylic acid can be treated with diphenylphosphoryl azide (DPPA) in an aprotic solvent such as methylene chloride in the presence of a base. In the next step of the reaction procedure, acyl azide (3) is converted to the corresponding isoquinolone (4) as shown in the reaction scheme. In that case, the acyl azide is heated to a temperature of about 190 ° C. in a high boiling solvent such as diphenylmethane. This reaction is general and results in substituted isoquinolones in moderate to high yield from the corresponding cinnamic acid derivatives. It should be noted that the cinnamic acid derivatives are commercially available or can be obtained directly from the corresponding benzaldehyde (1) derivative by condensing with malonic acid or its derivatives and using the Wittig reaction. The intermediate isoquinolone (4) can be treated with phosphorus oxychloride and converted to the corresponding 1-chloroisoquinoline. This reaction is common and can be applied to any isoquinolone described herein to convert a hydroxy substituent to the corresponding chloro compound.

  An alternative method for synthesizing isoquinoline ring systems is the Pomeranz-Fritsh method. This general method is described below. The process begins with the conversion of a benzaldehyde derivative (1) to a functionalized imine (2). The imine is then converted to an isoquinoline ring system by acid treatment at an elevated temperature.

上記のこのイソキノリン合成法は一般的であり、C8位が置換されているイソキノリン中間体を得るのに特に有用であることに留意すべきである(注：上記反応式の中間体(3)において、イソキノリン環のC8位を置換基R₆で置換する)。中間体イソキノリン(3)は、示した2工程法において対応する1-クロロキノリン (5)に変換することができる。この手順の第1工程は、ジクロロメタンのような非プロトン性溶媒中、イソキノリン (3)をメタクロロ過安息香酸で処理することによるイソキノリン N-オキシド(4)の形成である。中間体(4)を、還流クロロホルム中、オキシ塩化リンで処理することにより対応する1-クロロイソキノリンに変換することができる。

It should be noted that this isoquinoline synthesis method described above is general and is particularly useful for obtaining isoquinoline intermediates substituted at the C8 position (Note: in intermediate (3) of the above reaction scheme) And the C8 position of the isoquinoline ring is substituted with the substituent R ₆ ). The intermediate isoquinoline (3) can be converted to the corresponding 1-chloroquinoline (5) in the indicated two-step process. The first step in this procedure is the formation of isoquinoline N-oxide (4) by treating isoquinoline (3) with metachloroperbenzoic acid in an aprotic solvent such as dichloromethane. Intermediate (4) can be converted to the corresponding 1-chloroisoquinoline by treatment with phosphorus oxychloride in refluxing chloroform.

  Another method for synthesizing the isoquinoline ring system is shown below.

この方法では、オルソアルキルベンズアミド誘導体(1)を低温でTHFのような溶媒中、tert-ブチルリチウムのような強塩基で処理する。次に、この反応混合物にニトリル誘導体を加え、(1)の脱プロトンから誘導したアニオンとさらに反応させて(2)の形成を得る。この反応は一般的であり、置換イソキノリンの形成に用いることができる。上記反応式の中間体(2)は、本明細書に記載の方法により対応する1-クロロイソキノリンに変換することができる。イソキノリンを合成するためのさらなる方法を以下に示す。tert-ブチルリチウムを用いる中間体(1)の脱プロトン化は上記している。しかしながら、本発明の方法において、該中間体アニオンをエステルで捕捉し、下記のごとく中間体(2)を形成させる。次の反応において、ケトン(2)を上昇した温度で酢酸アンモニウムと縮合させてキノロン(3)を形成させる。この反応は一般的であり、置換されたイソキノロンの構築に応用することができ、次いで、本明細書に記載のごとく対応する1-クロロイソキノリンに変換することができる。

In this method, the orthoalkylbenzamide derivative (1) is treated with a strong base such as tert-butyllithium in a solvent such as THF at a low temperature. A nitrile derivative is then added to the reaction mixture and further reacted with an anion derived from the deprotonation of (1) to yield the formation of (2). This reaction is common and can be used to form substituted isoquinolines. Intermediate (2) in the above reaction scheme can be converted to the corresponding 1-chloroisoquinoline by the methods described herein. Additional methods for synthesizing isoquinolines are shown below. Deprotonation of intermediate (1) using tert-butyllithium is described above. However, in the method of the present invention, the intermediate anion is captured with an ester to form intermediate (2) as follows. In the next reaction, ketone (2) is condensed with ammonium acetate at an elevated temperature to form quinolone (3). This reaction is general and can be applied to the construction of substituted isoquinolones and then converted to the corresponding 1-chloroisoquinoline as described herein.

得られたケトアミン(2)を上昇した温度で酢酸アンモニウムと縮合させて対応するイソキノリンに変換することができる。この方法は一般的であり、置換されたイソキノリンの合成に用いることができる。該イソキノリンは、本明細書に記載の方法により対応する1-クロロイソキノリンに変換することができる。 A further method for constructing isoquinoline is shown in the following reaction scheme. In the first step of this procedure, an ortho-alkylarylimine derivative such as (1) is subjected to deprotonation conditions (sec-butyllithium, THF) and then the resulting anion is converted to an activated carboxylic acid derivative such as Weinreb amide. Damping by addition of.

The resulting ketoamine (2) can be converted to the corresponding isoquinoline by condensation with ammonium acetate at an elevated temperature. This method is general and can be used for the synthesis of substituted isoquinolines. The isoquinoline can be converted to the corresponding 1-chloroisoquinoline by the methods described herein.

  [4 + 2] cycloaddition reactions can also be used to construct functionalized isoquinoline ring systems. For example, as described below, a functionalized isoquinolone (3) is obtained using a vinyl isocyanate (1) in a cycloaddition reaction with a benzine precursor (2). The isoquinoline can be incorporated into a compound of formula I using the methods described herein.

  The isoquinolines incorporated into the compounds of formula I described herein can be further functionalized. It will be apparent to those skilled in the art that further functionalization of the heterocycle can be performed before or after incorporation of these functionalities into the compound of Formula I. The following reaction equation illustrates this point.

例えば、下記反応式は、アルコキシドが室温で誘導されるアルコール溶媒中のナトリウムまたはカリウムアルコキシド種による(eq.1)の(1)の処理による1-クロロ-6-フルオロ-イソキノリンの対応する1-クロロ-6-アルコキシ-イソキノリン種への変換を示す。ある場合には、反応物を加熱して反応を完結させる必要があるかもしれない。方程式1の中間体(2)を本明細書に記載のごとく式Iの化合物に組み込むことができる。方程式2に示すように、類似の反応は、6-フルオロ-イソキノリン要素が大環に組み込まれている基質上に行うこともできる。そこで、アルコキシドが誘導されるアルコール溶媒中のナトリウムまたはカリウムアルコキシド種による方程式2の中間体2の処理により式1の化合物を得ることができる。

For example, the following reaction scheme shows the corresponding 1-chloro-6-fluoro-isoquinoline by treatment of (eq. 1) (1) with a sodium or potassium alkoxide species in an alcohol solvent from which the alkoxide is derived at room temperature. Shows conversion to the chloro-6-alkoxy-isoquinoline species. In some cases, it may be necessary to heat the reactants to complete the reaction. Intermediate (2) of equation 1 can be incorporated into compounds of formula I as described herein. As shown in Equation 2, a similar reaction can also be performed on a substrate in which a 6-fluoro-isoquinoline element is incorporated into the macrocycle. Thus, the compound of formula 1 can be obtained by treatment of intermediate 2 of equation 2 with sodium or potassium alkoxide species in an alcohol solvent from which the alkoxide is derived.

  The reaction scheme below provides a general example for modifying isoquinoline as described herein by using a palladium mediated coupling reaction. The coupling can be used to functionalize the isoquinoline at each position of the ring system when the ring is appropriately activated or functionalized with chloride, for example as shown in the reaction scheme. This procedure can be started with 1-chloroisoquinoline (1) and converted to the corresponding N-oxide (2) by treatment with metachloroperbenzoic acid. The intermediate (2) can be converted to the corresponding 1,3-dichloroisoquinoline (3) by treatment with phosphorus oxychloride in chloroform. Intermediate (3) can be coupled with N-Boc-4-hydroxyproline by the methods described herein to give intermediate (5) as shown in the reaction scheme. Intermediate (5) can be Suzuki coupled with an arylboronic acid in the presence of a palladium reagent and a base and in a solvent such as THF or toluene or DMF to give a C3-arylisoquinoline intermediate (6). Heteroaryl boronic acids can also be used in this Pd-mediated coupling reaction to give C3-heteroarylisoquinolines. Intermediate (6) can be converted to the final compound of formula I by the methods described herein.

  Palladium-mediated coupling with heteroaryl-based aryl or heteroaryl elements can also be used in subsequent synthetic steps in the construction of compounds of formula I as follows. Therefore, the macrocyclic tripeptide acylsulfonamide intermediate (1) was coupled with 1-chloro-3-bromoisoquinoline (2) using the alkoxide substitution method of the heteroarylhalo moiety described above to obtain the intermediate (3 ) The coupling of (1) and (2) can be performed in the presence of a catalyst such as lanthanum chloride. The isoquinoline ring system of intermediate (3) is further coupled in a Suzuki coupling (Method 1: in a solvent such as DMF, in the presence of a palladium catalyst such as palladium tetrakis (triphenylphosphine) and a base such as cesium carbonate ( 3) heteroaryl or arylboronic acid), or Stille coupling (Method 2: heteroaryl in a solvent such as toluene in the presence of a palladium catalyst such as palladium tetrakis (triphenylphosphine) Or an aryltin derivative) can be functionalized.

  The invention also provides a composition comprising a compound of the invention, or a pharmaceutically acceptable enantiomer, diastereomer, salt, solvate, or prodrug thereof, and a pharmaceutically acceptable carrier. A pharmaceutical composition of the invention comprises a therapeutically effective amount of a compound of the invention, or a pharmaceutically acceptable enantiomer, diastereomer, salt, solvate, or prodrug thereof, and a pharmaceutically acceptable carrier, For example, an excipient or vehicle diluent.

  In such compositions, the active ingredient or compound typically comprises from 0.1% to 99.9% by weight of the composition, often from about 5 to 95% by weight of the composition.

  Thus, in one aspect of the invention, a composition comprising a compound of Formula 1 and a pharmaceutically acceptable carrier is provided. Preferably, the composition further comprises a compound having anti-HCV activity. As used herein, the term “anti-HCV activity” means that the compound is an HCV metalloprotease, HCV serine protease, HCV polymerase, HCV helicase, HCV NS4B protein, HCV entry, HCV in order to treat HCV infection. Means effective to inhibit the function of a target selected from the group consisting of assembly, HCV release, HCV NS5A protein, IMPDH, and nucleoside analogues. Often, other compounds with anti-HCV activity are effective in inhibiting the function of targets in the HCV life cycle other than the HCV NS3 protease protein.

  In certain preferred aspects, the compound having anti-HCV activity is interferon. Preferably, the interferon is selected from the group consisting of interferon alpha 2B, polyethylene glycol-added interferon alpha, consensus interferon, interferon alpha 2A, lymphoblastiod interferon tau.

  In another aspect of the present invention, the compound having anti-HCV activity is interleukin 2, interleukin 6, interleukin 12, a compound that enhances expression of type 1 helper T cell reaction, interfering RNA, antisense RNA, Imiqimod, Selected from the group consisting of ribavirin, inosine 5′-monophosphate dehydrogenase inhibitor, amantadine, and rimantadine.

  In certain preferred aspects of the invention, the composition comprises a compound of the invention, interferon, and ribavirin.

  In another preferred aspect of the invention, the compound having anti-HCV activity is a small molecule compound. As used herein, the term “small molecule compound” means a compound having a molecular weight of 1,500 daltons or less, and preferably a molecular weight of 1000 daltons or less. Preferably, the small molecule compound is HCV metalloprotease, HCV serine protease, HCV polymerase, HCV helicase, HCV NS4B protein, HCV entry, HCV assembly, HCV release, HCV NS5A protein, inosine to treat HCV infection It is effective to inhibit the function of a target selected from the group consisting of monophosphate dehydrogenase (“IMPDH”) and nucleoside analogues.

  Examples of certain HCV inhibitor compounds that can be administered with the compounds of the present invention include those disclosed in the following publications: WO 02/04425 A2 (published 17 January 2002), WO 03/007945 A1 (January 30, 2003), WO 03/010141 A2 (published February 6, 2003), WO 03/010142 A2 (published February 6, 2003), WO 03/010143 A1 (February 6, 2003) WO 03/000254 A1 (published on January 3, 2003), WO 01/32153 A2 (published on May 10, 2001), WO 00/06529 (published on February 10, 2000), WO 00 / 18231 (released April 6, 2000), WO 00/10573 (released March 2, 2000), WO 00/13708 (released March 16, 2000), WO 01/85172 A1 (November 2001) Published on 15th), WO 03/037893 A1 (published on May 8, 2003), WO 03/037894 A1 (published on May 8, 2003), WO 03/037895 A1 (published on May 8, 2003), WO 02/100851 A2 (published on December 19, 2002), WO 02/100846 A1 (published on December 19, 2002), EP 1256628 A2 (published on November 13, 2002), WO 99/01582 (1999) Published January 14), WO 00/09543 (published February 24, 2000).

  Table 1 below shows examples of some compounds that can be administered with the compounds of the present invention. The compounds of the present invention can be administered together with other anti-HCV active compounds, together or separately in combination therapy, or by mixing the compounds into the composition.

table 1

  The pharmaceutical composition of the present invention could be administered orally, parenterally or via an implantable reservoir. Oral administration or administration by injection is preferred. In some cases, the pH of the formulation may be adjusted with pharmaceutically acceptable acids, bases, or buffers to enhance the stability of the formulated compound or its delivery form. The term parenteral as used herein includes subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, and intralesional injection or infusion techniques.

  For oral administration, the pharmaceutical composition of the present invention could be administered in any orally acceptable dosage form including, but not limited to, capsules, tablets, and aqueous suspensions or solutions. In the case of tablets for oral use, carriers that are commonly used include lactose and corn starch. Typically, a lubricant such as magnesium stearate is also used. Diluents useful for oral administration in a capsule form include lactose and dried corn starch. When aqueous suspensions are administered orally, the active ingredient is mixed with emulsifying and suspending agents. If desired, certain sweetening and / or flavoring and / or coloring agents could be included. Other suitable carriers for the above compositions are described in standard pharmaceutical textbooks such as “Remington's Pharmaceutical Sciences”, 19th Edition, Mack Publishing Company, Easton, Penn., 1995.

  The pharmaceutical composition can be manufactured by known methods using well known and readily available ingredients. The compositions of the present invention could be formulated to release the active ingredient rapidly, sustained or delayed after administration to a patient using methods well known in the art. For preparing the compositions of the invention, the active ingredient is usually mixed with the carrier, diluted with a carrier, or encapsulated in a carrier that may be in the form of a capsule, sachet, paper, or other container. Like. When a carrier is used as a diluent, the carrier may be a solid, semi-solid, or liquid material that acts as a vehicle, excipient or medium for the active ingredient. That is, the composition comprises tablets, pills, powders, beadlets, lozenges, sachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solid or in a liquid medium), It may be in the form of soft and hard gelatin capsules, suppositories, sterile injectable solutions, sterile packaged powders and the like. Further details regarding the design and manufacture of suitable delivery forms for the pharmaceutical compositions of the present invention are known to those skilled in the art.

  A dosage level of about 0.01 to about 1000 milligrams / kilogram (“mg / kg”) body weight / day, preferably about 0.5 to about 250 mg / kg body weight / day, of the compound of the present invention is used alone for the prevention and treatment of HCV-mediated diseases. Typical for therapy. Typically, the pharmaceutical compositions of this invention will be administered from about 1 to about 5 times per day, or as a continuous infusion. Such administration can be used as a chronic or acute therapy. The amount of active ingredient that can be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated and the particular mode of administration.

  As those skilled in the art will appreciate, lower or higher doses may be necessary. The specific dose and treatment plan for any particular patient will depend on the activity, age, weight, general health, sex, diet, frequency of administration, excretion rate, combination of drugs, severity and course of infection, It will depend on a variety of factors, including the disposition of the infection and the judgment of the treating physician. Generally, treatment is initiated with a dosage that is substantially less than the optimum amount of the peptide. Thereafter, the dosage is increased by small increments until the optimum effect under circumstances is reached. In general, it is most desirable to administer the compounds at a concentration level that will provide an antiviral effect without causing any harmful or untoward side effects.

  Where a composition of the invention comprises a combination of a compound of the invention and one or more additional therapeutic or prophylactic agents, the compound and the additional agent are usually about 10 to 10% of the dose normally administered in a monotherapy regimen. It is present at a dose level of 100%, more preferably about 10-80%.

  When these compounds, or pharmaceutically acceptable enantiomers, diastereomers, salts, solvates, or prodrugs thereof, are formulated with a pharmaceutically acceptable carrier, the resulting composition is fed in vivo. It could be administered to an animal, such as a human, to inhibit HCV NS3 protease or prevent HCV viral infection.

  Accordingly, another aspect of the present invention provides a method for inhibiting HCV NS3 protease activity in a patient by administering a compound of the present invention or a pharmaceutically acceptable enantiomer, diastereomer, salt, or solvate thereof. It is to be.

  One aspect of the present invention provides a patient's HCV comprising administering to the patient a therapeutically effective amount of a compound of the present invention, or a pharmaceutically acceptable enantiomer, diastereomer, solvate, prodrug, or salt thereof. It is to provide a method of treating an infection.

  Preferably, the method of administering the compound is effective to inhibit the function of HCV NS3 protease protein. In a preferred aspect, the method further comprises administering another compound (described above) having anti-HCV activity before, after, or simultaneously with the compound of the present invention.

  The compounds of the present invention can also be used as experimental reagents. The compounds may be beneficial in designing viral replication assays to further increase knowledge of the mechanisms of HCV disease, validation of animal assay systems, and providing research tools for structural biological testing. Furthermore, the compounds of the invention are useful for establishing or determining the binding site of other antiviral compounds, for example by competitive inhibition.

  The compounds of the present invention are used to treat or prevent viral contamination of substances and to contact such substances such as blood, tissue, surgical instruments and clothing, laboratory instruments and clothing, and blood collection or transfusion devices and substances. It may also reduce the risk of viral infections in experiments or medical workers or patients.

  Furthermore, the compounds and compositions of the present invention can be used in the manufacture of a medicament for treating HCV infection in a patient.

  The present invention relates to an alkyl ester enantiomer characterized in that the contacting is carried out in the presence of a buffer comprising contacting the mixture with an enzyme effective to selectively promote hydrolysis of one of the enantiomers There is also provided a method for dividing the mixture. Such a mixture of alkyl ester enantiomers can result from the preparation of chemical precursor B above.

［式中、R₂₅はアミノ保護基、例えばカルバメート、例えばBOCイミンもしくはアミドなどであり、R₂₆はC_1-10アルキル、C_6-14アリール、C_7-16アルキルアリール、C_3-7シクロアルキル、またはC_3-10アルキルシクロアルキルからなる群から選ばれる。］ Preferably, the alkyl ester has the formula:

[Wherein R ₂₅ is an amino protecting group such as a carbamate such as BOC imine or amide, and R ₂₆ is C _1-10 alkyl, C _6-14 aryl, C _7-16 alkylaryl, C _3-7 cyclo It is selected from the group consisting of alkyl or C _3-10 alkylcycloalkyl. ]

The particular buffer used in the method of the present invention is not critical. Typically, the buffer will have a pKa of ± 1 at which the hydrolysis takes place, for example, a pH of about 7.0-11. That is, when hydrolysis is performed at, for example, pH 8.0, a buffer having a pKa of about 7.0 to 9.0 can be used. Preferred buffering agents are phosphates, borates, and carbonates. Examples of suitable buffering agents include 2-[(2-amino-2-oxoethyl) amino] ethanesulfonic acid,
N- (2-acetamido) -2-iminodiacetic acid, n, n-bis (2-hydroxyethyl) glycine,
1,3-bis [tris (hydroxymethyl) methylamino] propane, o-boric acid, carbonic acid,
2- (cyclohexylamino) ethanesulfonic acid, diethylmalonic acid, glycylglycine,
N-2-hydroxyethylpiperazine-N'-2-ethane-sulfonic acid (HEPES),
N-2-hydroxyethylpiperazine-N'-3-propane-sulfonic acid, imidazole,
3- (N-morpholino) propanesulfonic acid, o-phosphoric acid,
Piperazine-N-N'-bis (2-ethanesulfonic acid),
Piperazine-1,4-bis (2-hydroxypropanesulfonic acid),
3- [tris (hydroxymethyl) methyl] aminopropanesulfonic acid,
2- [tris (hydroxymethyl) methyl] aminoethanesulfonic acid,
N- [tris (hydroxymethyl) methyl] glycine and tris (hydroxymethyl) aminomethane are included.

  Enzymes effective to selectively hydrolyze the desired alkyl ester enantiomer can be used in accordance with the present invention. Proteases, lipases, and esterases are often used for hydrolysis of carboxylic ester bonds. Enzymes suitable for use in accordance with the present invention may be used in free form, in liquid or dry form, or immobilized on a support by physical adsorption or incorporation, regardless of origin or purity. Examples of lipases include lipases from Candida rugosa, malt, porcine pancreas, Rhizopus arrhizus, Candida antarctica, Mucor miehei, Aspergillus niger, Pseudomonas species, Candida lipolytica, Humicola langinosa, and Mucor javanicus. Examples of esterases include porcine liver esterase (PLE), horse liver esterase (HLE), acetylcholinesterase (ACE), cholesterol esterase, and esterase derived from Bacillus subtilis (carboxyl esterase NP). Proteases are generally defined as hydrolases that act on peptide bonds as their natural substrate, but ester bonds are much weaker than peptide (amide) bonds and can cleave carboxylic acid ester bonds. Examples of protease include proteases derived from α-chymotrypsin, thermolysin, papain, Aspergillus sp., Bacillus sp., Rhizopus sp., Penicillum sp., Streptomyces griseus.

  In the ester of formula VIII, the chiral center of the acid moiety is fully substituted. Since their natural substrates (eg natural amino acids) usually contain at least one α-hydrogen, this means a different target of the hydrolase. Quite surprisingly, it has been found according to the present invention that certain proteases are highly effective in the enantioselective hydrolysis of ester VIII. Preferred proteases include proteases from strains selected from the group consisting of Bacillus globigii, Bacillus licheniformis, Bacillus halodurans, Bacillus clausii, and Aspergillus oryzae. Examples of suitable enzymes include Alcalase® (subtilisin, alkaline protease, Novozymes North America Inc., Franklington, NC), Savinase® (subtilisin, alkaline protease, Novozymes), ChiroCLEC® Included are those selected from the group consisting of BL (subtilisin, alkaline protease, Altus Biologics, Inc.,), ProteaseN (subtilisin, alkaline protease, Amano), and Flavorzyme® (fungal protease / peptidase, Novozymes). Further details of such enzymes are known in the art. Such enzymes are commercially available. These enzymes can be used alone or as a mixture thereof.

  The hydrolysis of the ester results in the release of alcohol and carboxylic acid and can significantly reduce the pH in the absence of a buffer. According to the present invention, it has been found that the inclusion of a buffer in the reaction mixture can stabilize the pH during the hydrolysis process, contributing to higher enzyme activity and stability.

  Furthermore, according to the present invention, the hydrolysis can be carried out at an elevated temperature, preferably about 30-60 ° C., for a short period of time, preferably about 7 days or less, more preferably about 2 hours to 3 days.

  The following specific examples illustrate the synthesis of the compounds of the present invention and are not intended to limit the scope of the claims. The method could be adapted to variations to produce the compounds included in the invention not specifically disclosed. In addition, variations in the process for preparing the compounds in somewhat different ways will be apparent to those skilled in the art.

Chemical abbreviations commonly used to identify the compounds described in this invention include: Bn: benzyl; Boc: tert-butyloxycarbonyl {Me ₃ COC (O)}; BSA: bovine serum albumin; CDI: carbonyl DBU: 1,8-diazabicyclo [5.4.0] -undec-7-ene; CH ₂ Cl ₂ = DCM: methylene chloride; TBME: tert-butyl methyl ether; DEAD: diethyl azodicarboxylate; DIAD: Diisopropyl azodicarboxylate; DIEA: diisopropylethylamine; DIPEA: diisopropylethylamine; 4-DMAP: 4-dimethylaminopyridine; DCC: 1,3-bicyclohexylcarbodiimide; DMF: dimethylformamide; DMSO: dimethylsulfoxide; DPPA: diphenylphosphoryl azido; Et: ethyl; EtOH: ethanol; EtOAc: ethyl acetate; Et ₂ O: diethyl ether; Grubb catalyst: bis (tri-cyclohexylene Phosphine) benzylideneruthenium (IV) dichloride; Grubb second generation catalyst: tricyclohexylphosphine [1,3-bis (2,4,6-trimethylphenyl) -4,5-dihydroimidazol-2-ylidene] [benzylidene] ruthenium (IV) Dichloride; HATU: [O- (7-azabenzotriazol-1-yl) -N, N, N ′, N′-tetramethyluronium hexafluorophosphate; HBTU: [O- (1H-benzotriazole -1-yl) -N, N, N ′, N′-tetramethyluronium hexafluorophosphate; HOBT, 1-hydroxybenzotriazole; HOAT, 1-hydroxy-7-azabenzotriazole; HPLC: high performance liquid chromatography; MS: mass spectrometry; Me: methyl; MeOH: methanol; NMM: N-methylmorpholine; NMP: N-methylpyrrolidine; Pr: propyl; PPA: polyphosphoric acid; TBAF: tetra-n-butylammonium fluoride; 2-DCE or DCE: 1,2 -Dichloroethane; TFA: trifluoroacetic acid; THF: tetrahydrofuran.

Unless otherwise noted, solution percentages are expressed in weight / volume relationships and solution ratios are expressed in volume / volume. Nuclear magnetic resonance (NMR) spectra were recorded using a Bruker 300, 400, or 500 megahertz (MHz) spectrophotometer; chemical shifts (δ) were reported in parts per million. Flash chromatography was performed using silica gel (SiO ₂ ), which will be apparent to those skilled in the art. All liquid chromatography (LC) data is recorded with a Shimadzu LC-10AS liquid chromatograph using an SPD-10AV UV-Vis detector, and mass spectrometry (MS) data using the Micromass Platform for LC in an electrospray method. Measured (ES +).

Unless otherwise noted, each compound was analyzed by LC / MS using one of three methodologies with the following conditions.
Column: Method A) -YMC Xterra ODS S7 micrometer (“μm”) 3.0 × 50 millimeters (“mm”), ie diameter 3.0 mm, height 50 mm.
Gradient: 100% solvent A / 0% solvent B to 0% solvent A / 100% solvent B
Gradient time: 4min (A)
Holding time: 1min (A); 2min
Flow rate: 4mL / min
Detector wavelength: 220 nanometers (“nm”)
Solvent A: 10% MeOH / 90% H ₂ O / 0.1% TFA
Solvent B: 10% H ₂ O / 90% MeOH / 0.1% TFA.
Column: Method B) -YMC Xterra ODS S7 3.0x50mm
Gradient: 100% solvent A / 0% solvent B to 0% solvent A / 100% solvent B
Gradient time: 2 min (minutes)
Holding time: 1min
Flow rate: 5mL / min
Detector wavelength: 220nm
Solvent A: 10% MeOH / 90% H ₂ O / 0.1% TFA
Solvent B: 10% H ₂ O / 90% MeOH / 0.1% TFA.
Column: Method D) -YMC Xterra C18 5μM 3.0x50mm
Gradient: 100% solvent A / 0% solvent B to 0% solvent A / 100% solvent B
Gradient time: 3min
Holding time: 1min
Flow rate: 5mL / min
Detector wavelength: 220nm
Solvent A: 10% MeOH / 90% H ₂ O / 0.1% TFA
Solvent B: 10% H ₂ O / 90% MeOH / 0.1% TFA.

Unless otherwise stated, preparative HPLC was performed under the following conditions.
Gradient time: 10min
Holding time: 2min
Flow rate: 25mL / min
Detector wavelength: 220 nM
Solvent A: 10% MeOH / 90% H ₂ O / 0.1% TFA
Solvent B: 10% H ₂ O / 90% MeOH / 0.1% TFA

The pure fractions are mixed and brought to pH up to 7 using 0.1N NaOH. The solution is concentrated under reduced pressure and then the pH is carefully lowered to pH 3-4 with 0.1N HCl. The mixture is immediately extracted with ethyl acetate (3x). The combined extracts are dried (MgSO ₄ ) and then concentrated under reduced pressure to give the purified product.

The compounds and chemical intermediates of the present invention described in the following examples were prepared according to the following methods.
Example 1 Preparation of a Typical Intermediate Preparation of a P2 Isoquinoline Intermediate for Incorporation into a Compound of Formula 1
Method A

工程1:

Process 1 :

To a solution of acetone (80 mL) in 3-methoxycinnamic acid (11.04 g, 62 mmol) and triethylamine (12.52 g, 124 mmol) was added ethyl chloroformate (about 1.5 eq) at 0 ° C. After stirring for 1 hour at this temperature, aqueous NaN ₃ (6.40 g, 100 mmol, in 35 mL H ₂ O; appropriate precautions must be taken when using sodium azide) is added dropwise and the reaction mixture is allowed to cool to ambient temperature. For 16 hours. Water (100 mL) was added to the mixture and volatiles were removed under reduced pressure. The resulting slurry was extracted with toluene (3 × 50 mL) and the combined organic layer was dried over MgSO ₄ . This dry solution was added dropwise to a solution of diphenylmethane (50 mL) and tributylamine (30 mL) heated to 190 ° C. The added toluene was distilled off. After complete addition, the reaction temperature was raised to 210 ° C. for 2 hours. After cooling, the precipitated product was collected by filtration, washed with hexane (2 × 50 mL) and then dried to give the desired product as a white solid (5.53 g, 51%) (Nicolas Briet at el, Tetrahedron 2002, 5761-5766).
LC-MS (retention time: 0.82 min, method B), MS m / z 176 (M + + H).

An alternative to the above uses diphenylphosphoryl azide to convert the carboxylic acid to the corresponding acyl azide. The acid is then converted to the corresponding quinolone in a one pot process. The process is described below for the preparation of 4-methyl-2H-isoquinolin-1-one from 3-phenyl-but-2-enoic acid.
A solution of 3-phenyl-but-2-enoic acid (16.2 g), diphenylphosphoryl azide (27.5 g), and triethylamine (10.1 g) in benzene (100 mL) was stirred for 1 hour. After filtration through a silica gel plug washed with benzene and concentration, the residue was dissolved in diphenylmethane (80 mL) and refluxed for 3 hours. After cooling to room temperature, the solid was collected on a plug washed with benzene and then dried to give 10 g (63%) of the desired 4-methyl-2H-isoquinolin-1-one as a solid. ¹ H NMR (400 MHz, CD ₃ OD) δ ppm 2.30 (s, 3H), 7.00 (s, 1H), 7.54 (m, 1H), 7.77 (m, 2H), 8.33 (d, J = 7.34Hz, 1H ).
Process 2 :

6-Methoxy-2H-isoquinolin-1-one (5.0 g, 28.4 mmol) in POCl ₃ (10 mL) was gently heated to reflux for 3 hours and evaporated under reduced pressure (Nicolas Briet et al, Tetrahedron, 2002, 5761-5766). The residue was poured into ice water (20 mL) and neutralized to pH 10 with 10 M NaOH. Extracted with CHCl ₃ . The organic layer was washed with brine, dried over MgSO ₄ , filtered and evaporated. The residue was purified by flash chromatography (1: 1 hexane-EtOAc) to give 4.41 g (80%) of the desired product as a white solid.
¹ H NMR (CD ₃ OD) δ 3.98 (s, 3H), 7.34-7.38 (m, 2H), 7.69 (d, J = 5.5Hz, 1H), 8.10 (d, J = 6.0Hz, 1H), 8.23 (d, J = 9.5Hz, 1H);
LC-MS (retention time: 1.42min, method B), MS m / z 194 (M + + H).
Process 3 :

To a solution of N-BOC-3- (R) -hydroxy-L-proline (892 mg, 3.89 mmol) in DMSO (40 mL) at ambient temperature was added potassium tert-butoxide (1.34 g, 12.0 mmol) in one portion. . The formed suspension was stirred at this temperature for 30 minutes and then cooled to 100 ° C. 1-Chloro-6-methoxy-isoquinoline (Example 11, Step 2) (785 mg, 4.05 mmol) was added in one portion as a solid and the final mixture was stirred for 12 hours at ambient temperature. Attenuated with ice-cold 5% citric acid (aq) and then extracted with EtOAC (100 mL). The aqueous phase was extracted again with EtOAC. The combined organic layers were each washed with 5% citric acid (aq) and brine, dried over MgSO ₄ and filtered. The filtrate was evaporated under reduced pressure to give 1.49 g (99%) of the desired product as an off-white foam. This material was used in the next step reaction as a crude material without further purification.
¹ H NMR (CD ₃ OD) δ 1.42, 1.44 (rotamers, 9H), 2.38-2.43 (m, 1H), 2.66-2.72 (m, 1H), 3.80-3.87 (m, 2H), 3.92 (s 3H), 4.44-4.52 (m, 1H), 5.73 (b, 1H), 7.16-7.18 (m, 2H), 7.24-7.25 (m, 1H), 7.87-7.88 (m, 1H), 8.07 (d , J = 8.5Hz, 1H);
LC-MS (retention time: 1.62min, method B), MS m / z 389 (M + + H).

  The following intermediates can be prepared as described herein and incorporated into compounds of Formula 1.

工程1:
修飾：15gの3-メトキシ-3-フェニル-アクリル酸を用い、250mgの生成物を得た (2%(収率))。
生成物:

Process 1 :
Modification : Using 15 g of 3-methoxy-3-phenyl-acrylic acid, 250 mg of product was obtained (2% (yield)).
Product :

¹H NMR (400MHz、CD₃COCD₃) δ ppm 3.85 (s、3H)、6.96 (s、1H)、7.54 (m、1H)、7.71 (m、1H)、7.86 (d、J=8.07Hz、1H)、8.31 (d、J=8.07Hz、1H)。
工程2：
修飾：200mgの4-メトキシ-2H-イソキノリン-1-オンを用い、150mgの生成物を得た(68%(収率))。
生成物:

¹ H NMR (400 MHz, CD ₃ COCD ₃ ) δ ppm 3.85 (s, 3H), 6.96 (s, 1H), 7.54 (m, 1H), 7.71 (m, 1H), 7.86 (d, J = 8.07Hz, 1H), 8.31 (d, J = 8.07 Hz, 1H).
Process 2 :
Modification: Using 200 mg 4-methoxy-2H-isoquinolin-1-one, 150 mg product was obtained (68% (yield)).
Product :

¹H NMR (400MHz、CDCl₃) δ ppm 4.05 (s、2H)、7.71 (m、1H)、7.72 (m、2H)、7.80 (s、1H)、8.23 (dd、J=18.71、7.70Hz、2 H)。
工程3：
修飾：122mgの1-クロロ-4-メトキシ-イソキノリンを用い、218mgの生成物を得た(89%(収率))。
生成物：

¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 4.05 (s, 2H), 7.71 (m, 1H), 7.72 (m, 2H), 7.80 (s, 1H), 8.23 (dd, J = 18.71, 7.70 Hz, 2H).
Process 3 :
Modification: 122 mg of 1-chloro-4-methoxy-isoquinoline was used to give 218 mg of product (89% (yield)).
Product :

MS：(M+Na)⁺ 411。

^{MS: (M + Na) +} 411.

  The following intermediates can be prepared as described herein and incorporated into compounds of Formula 1.

工程1：
修飾：20gの2-メチル桂皮酸を用い、14.3gの生成物を得た(72%(収率))。
生成物：

Process 1 :
Modification : Using 20 g of 2-methylcinnamic acid, 14.3 g of product was obtained (72% (yield)).
Product :

データ：¹H NMR (400MHz、CD₃OD) δ ppm 2.54 (s、1H)、6.69 (d、J=7.3Hz、1H)、7.23 (d、J=7.3Hz、1H)、7.39 (t、J=7.8Hz、1H)、7.50 (d、J=7.1Hz、1H)、8.30 (d、J=8.1Hz、1H)、11.62 (s、1H)；MS：(M+H)⁺ 160。
工程2：
修飾：14.4gの5-メチル-2H-イソキノリン-1-オンを用い、10.6gの生成物を得た(66%(収率))。
生成物：

Data: ¹ H NMR (400MHz, CD ₃ OD) δ ppm 2.54 (s, 1H), 6.69 (d, J = 7.3Hz, 1H), 7.23 (d, J = 7.3Hz, 1H), 7.39 (t, J = 7.8Hz, 1H), 7.50 ( d, J = 7.1Hz, 1H), 8.30 (d, J = 8.1Hz, 1H), 11.62 (s, 1H); MS: (M + H) + 160.
Process 2 :
Modification : 14.4 g of 5-methyl-2H-isoquinolin-1-one was used to give 10.6 g of product (66% (yield)).
Product :

データ：¹H NMR (400MHz、CDCl₃) δ ppm 2.67 (s、3H)、7.55 (m、2H)、7.70 (dd、J=5.9、1.0Hz、1H)、8.19 (m、1H)、8.28 (d、J=5.9Hz、1H)；MS：(M+H)⁺ 178。
工程3：
修飾：533mgの1-クロロ-5-メチル-イソキノリンを用い、1116mgの生成物を得た(100%(収率))。
生成物：

Data: ¹ H NMR (400MHz, CDCl ₃ ) δ ppm 2.67 (s, 3H), 7.55 (m, 2H), 7.70 (dd, J = 5.9, 1.0Hz, 1H), 8.19 (m, 1H), 8.28 ( d, J = 5.9Hz, 1H) ; MS: (M + H) + 178.
Process 3 :
Modification: 533 mg of 1-chloro-5-methyl-isoquinoline was used to give 1116 mg of product (100% (yield)).
Product :

データ：MS：(M+H)⁺ 373。
以下の中間体を本明細書に記載のごとく製造し、式1の化合物に組み込むことができる。

Data: MS: (M + H) + 373.
The following intermediates can be prepared as described herein and incorporated into compounds of Formula 1.

工程1：
修飾：10gの2-メトキシ桂皮酸を用い、5.3gの生成物を得た(53%(収率))。
生成物：

Process 1 :
Modification : Using 10 g of 2-methoxycinnamic acid, 5.3 g of product was obtained (53% (yield)).
Product :

データ：¹H NMR (400MHz、CD₃OD) δ ppm 3.95 (s、3H)、6.94 (d、J=7.3Hz、1H)、7.08 (d、J=8.1Hz、1H)、7.14 (d、J=7.3Hz、1H)、7.43 (t、J=8.1Hz、1H)、7.99 (d、J=8.1Hz、1H)、10.92 (s、1H)；MS：(M+H)⁺ 176.
工程2：
修飾：5.3gの5-メトキシ-2H-イソキノリン-1-オンを用い、5.38gの生成物を得た(92%(収率))。
生成物：

Data: ¹ H NMR (400MHz, CD ₃ OD) δ ppm 3.95 (s, 3H), 6.94 (d, J = 7.3Hz, 1H), 7.08 (d, J = 8.1Hz, 1H), 7.14 (d, J = 7.3Hz, 1H), 7.43 (t, J = 8.1Hz, 1H), 7.99 (d, J = 8.1Hz, 1H), 10.92 (s, 1H); MS: (M + H) ⁺ 176.
Process 2 :
Modification: Using 5.3 g of 5-methoxy-2H-isoquinolin-1-one, 5.38 g of product was obtained (92% (yield)).
Product :

データ：¹H NMR (400MHz、CDCl₃) δ ppm 4.01 (s、3H)、7.04 (d、J=7.8Hz、1H)、7.57 (t、J=8.1Hz、1H)、7.88 (d、J=8.6Hz、1H)、7.97 (d、J=5.9Hz、1H)、8.25 (d、J=5.9Hz、1H)；MS：(M+H)⁺ 194.
工程3：
修飾：581mgの1-クロロ-5-メトキシ-イソキノリンを用い、1163mgの生成物を得た(100%(収率))。
生成物：

Data: ¹ H NMR (400MHz, CDCl ₃ ) δ ppm 4.01 (s, 3H), 7.04 (d, J = 7.8Hz, 1H), 7.57 (t, J = 8.1Hz, 1H), 7.88 (d, J = 8.6Hz, 1H), 7.97 (d, J = 5.9Hz, 1H), 8.25 (d, J = 5.9Hz, 1H); MS: (M + H) ⁺ 194.
Process 3 :
Modification : 581 mg of 1-chloro-5-methoxy-isoquinoline was used to give 1163 mg of product (100% (yield)).
Product :

データ： MS：(M+H)⁺ 389.
以下の中間体を本明細書に記載のごとく製造し、式1の化合物に組み込むことができる。

Data: MS: (M + H) ⁺ 389.
The following intermediates can be prepared as described herein and incorporated into compounds of Formula 1.

工程1：
修飾：25gの2-クロロ桂皮酸を用い、14.6gの生成物を得た(59%(収率))。
生成物：

Process 1 :
Modification : 14.6 g of product was obtained using 25 g of 2-chlorocinnamic acid (59% (yield)).
Product :

データ：¹H NMR (400MHz、CD₃OD ) δ ppm 7.22 (d、J=7.3Hz、1H)、7.42 (t、J=7.8Hz、1H)、7.73 (d、J=7.8Hz、1H)、8.34 (d、J=8.1Hz、1H)、10.61 (s、1H)；MS：(M+H)⁺ 180.
工程2：
修飾：14.2gの5-クロロ-2H-イソキノリン-1-オンを用い、8.28gの生成物を得た(53%(収率))。
生成物：

Data: ¹ H NMR (400 MHz, CD ₃ OD) δ ppm 7.22 (d, J = 7.3 Hz, 1H), 7.42 (t, J = 7.8 Hz, 1H), 7.73 (d, J = 7.8 Hz, 1H), 8.34 (d, J = 8.1Hz, 1H), 10.61 (s, 1H); MS: (M + H) ⁺ 180.
Process 2 :
Modification : Using 14.2 g of 5-chloro-2H-isoquinolin-1-one, 8.28 g of product was obtained (53% (yield)).
Product :

データ：¹H NMR (400MHz、CDCl₃) δ ppm 7.60 (dd、J=8. 6、7.6Hz、1H)、7.83 (m、1H)、8.00 (d、J=5.9Hz、1H)、8.29 (dt、J=8.9、1.0Hz、1H)、8.38 (d、J=5.9Hz、1H)；MS：(M+H)⁺ 198.
工程3：
修飾：594mgの1,5-ジクロロ-イソキノリンを用い、1174mgの生成物を得た(100%(収率))。
生成物：

Data: ¹ H NMR (400MHz, CDCl ₃ ) δ ppm 7.60 (dd, J = 8.6, 7.6Hz, 1H), 7.83 (m, 1H), 8.00 (d, J = 5.9Hz, 1H), 8.29 ( dt, J = 8.9,1.0Hz, 1H) , 8.38 (d, J = 5.9Hz, 1H); MS: (M + H) + 198.
Process 3 :
Modification : Using 594 mg of 1,5-dichloro-isoquinoline, 1174 mg of product was obtained (100% (yield)).
Product :

データ：MS：(M+H)⁺ 393.

Data: MS: (M + H) ⁺ 393.

  The following intermediates can be prepared as described herein and incorporated into compounds of Formula 1.

工程1：
修飾：16.6gの2-フルオロ桂皮酸を用い、8.55gの生成物を得た(51%(収率))。
生成物：

Process 1 :
Modification : 8.55 g of product was obtained using 16.6 g of 2-fluorocinnamic acid (51% yield).
Product :

データ：¹H NMR (400MHz、CD₃COCD₃) δ ppm 6.62 (d、J=7.3Hz、1H)、7.32 (d、J=7.3Hz、1H)、7.47 (m、2H)、8.09 (m、1H).
工程2：
修飾：8.4gの5-フルオロ-2H-イソキノリン-1-オンを用い、7.5gの生成物を得た(80%(収率))。
生成物：

Data: ¹ H NMR (400 MHz, CD ₃ COCD ₃ ) δ ppm 6.62 (d, J = 7.3 Hz, 1H), 7.32 (d, J = 7.3 Hz, 1H), 7.47 (m, 2H), 8.09 (m, 1H).
Process 2 :
Modification : Using 8.4 g of 5-fluoro-2H-isoquinolin-1-one, 7.5 g of product was obtained (80% (yield)).
Product :

データ：¹H NMR (400MHz、CDCl₃) δ ppm 7.43 (ddd、J=9.7、7.8、0.9Hz、1H)、7.62 (td、J=8.2、5.4Hz、1H)、7.84 (d、J=5.6Hz、1H)、8.14 (d、J=8.6Hz、1H)、8.33 (d、J=5.9Hz、1H)；MS：(M+H)⁺ 182.
工程3：
修飾：203mgの1-クロロ-5-フルオロ-イソキノリンを用い、384mgの生成物を得た(90%(収率))。
生成物：

Data: ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 7.43 (ddd, J = 9.7, 7.8, 0.9 Hz, 1H), 7.62 (td, J = 8.2, 5.4 Hz, 1H), 7.84 (d, J = 5.6 Hz, 1H), 8.14 (d , J = 8.6Hz, 1H), 8.33 (d, J = 5.9Hz, 1H); MS: (M + H) + 182.
Process 3 :
Modification : 203 mg of 1-chloro-5-fluoro-isoquinoline was used to give 384 mg of product (90% (yield)).
Product :

データ：¹H NMR (400MHz、CD₃SOCD₃) δ ppm 1.34、1.36 (2s、9H、回転異性体)、2.35 (m、1H)、2.61 (m、1H)、3.65 (d、J=12.23Hz、1H)、3.80 (m、1H)、4.35 (m、1H)、5.70 (s、1H)、7.48 (d、J=6.11Hz、1H)、7.63 (m、2H)、7.99 (m、1H)、8.10 (d、J=5.87Hz、1H)；MS：(M+Na)⁺ 399.

Data: ¹ H NMR (400 MHz, CD ₃ SOCD ₃ ) δ ppm 1.34, 1.36 (2s, 9H, rotamer), 2.35 (m, 1H), 2.61 (m, 1H), 3.65 (d, J = 12.23Hz , 1H), 3.80 (m, 1H), 4.35 (m, 1H), 5.70 (s, 1H), 7.48 (d, J = 6.11Hz, 1H), 7.63 (m, 2H), 7.99 (m, 1H) 8.10 (d, J = 5.87 Hz, 1H); MS: (M + Na) ⁺ 399.

  The following intermediates can be prepared as described herein and incorporated into compounds of Formula 1.

工程1：
修飾：16.6gの4-フルオロ桂皮酸を用い、8.2gの生成物を得た(49%(収率))。
生成物：

Process 1 :
Modification : Using 16.6 g of 4-fluorocinnamic acid, 8.2 g of product was obtained (49% (yield)).
Product :

データ：¹H NMR (400MHz、CD₃COCD₃) δ ppm 6.57 (d、J=7.09Hz、1H)、7.21 (d、J=7.09Hz、1H)、7.50 (m、1H)、7.72 (dd、J=8.68、5.26Hz、1H)、7.90 (dd、J=9.54、2.93Hz、1H).
工程2：
修飾：8.15gの7-フルオロ-2H-イソキノリン-1-オンを用い、7.6gの生成物を得た(84%(収率))。
生成物：

Data: ¹ H NMR (400 MHz, CD ₃ COCD ₃ ) δ ppm 6.57 (d, J = 7.09Hz, 1H), 7.21 (d, J = 7.09Hz, 1H), 7.50 (m, 1H), 7.72 (dd, J = 8.68, 5.26Hz, 1H), 7.90 (dd, J = 9.54, 2.93Hz, 1H).
Process 2 :
Modification : 8.15 g of 7-fluoro-2H-isoquinolin-1-one was used to give 7.6 g of product (84% yield).
Product :

データ：¹H NMR (400MHz、CDCl₃) δ ppm 7.52 (td、J=8.6、2.6Hz、1H)、7.59 (d、J=5.6Hz、1H)、7.86 (dd、J=9.1、5.4Hz、1H)、7.95 (dd、J=9.5、2.5Hz、1H)、8.26 (d、J=5.6Hz、1H)；MS：(M+H)⁺ 182.
工程3：
修飾：191mgの1-クロロ-7-フルオロ-イソキノリンを用い、350mgの生成物を得た(93%(収率))。
生成物：

Data: ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 7.52 (td, J = 8.6, 2.6 Hz, 1H), 7.59 (d, J = 5.6 Hz, 1H), 7.86 (dd, J = 9.1, 5.4 Hz, 1H), 7.95 (dd, J = 9.5,2.5Hz, 1H), 8.26 (d, J = 5.6Hz, 1H); MS: (M + H) + 182.
Process 3 :
Modification : 191 mg of 1-chloro-7-fluoro-isoquinoline was used to give 350 mg of product (93% (yield)).
Product :

データ：MS：(M+Na)⁺ 399.

Data: MS: (M + Na) ⁺ 399.

  The following intermediates can be prepared as described herein and incorporated into compounds of Formula 1.

工程1：
修飾：9.13gの4-クロロ桂皮酸を用い、4gの生成物を得た(44%(収率))。
生成物：

Process 1 :
Modification : Using 9.13 g of 4-chlorocinnamic acid, 4 g of product was obtained (44% (yield)).
Product :

データ：¹H NMR (400MHz、CD₃SOCD₃) δ ppm 6.58 (d、J=7.1Hz、1H)、7.20 (dd、J=7.1、5.9Hz、1H)、7.72 (m、2H)、8.10 (m、1H).
工程2：
修飾：3.5gの7-クロロ-2H-イソキノリン-1-オンを用い、2.8gの生成物を得た(72%(収率))。
生成物：

Data: ¹ H NMR (400 MHz, CD ₃ SOCD ₃ ) δ ppm 6.58 (d, J = 7.1 Hz, 1H), 7.20 (dd, J = 7.1, 5.9 Hz, 1H), 7.72 (m, 2H), 8.10 ( m, 1H).
Process 2 :
Modification : Using 3.5 g of 7-chloro-2H-isoquinolin-1-one, 2.8 g of product was obtained (72% (yield)).
Product :

データ：¹H NMR (500MHz、CDCl₃) δ ppm 7.59 (d、J=5.5Hz、1H)、7.69 (dd、J=8.9、2.1Hz、1H)、7.80 (d、J=8.6Hz、1H)、8.29 (d、J=5.5Hz、1H)、8.34 (s、1H)；MS：(M+H)⁺ 198.
工程3：
修飾：208mgの1,7-ジクロロ-イソキノリンを用い、350mgの生成物を得た(89%(収率))。
生成物：

Data: ¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 7.59 (d, J = 5.5 Hz, 1H), 7.69 (dd, J = 8.9, 2.1 Hz, 1H), 7.80 (d, J = 8.6 Hz, 1H) 8.29 (d, J = 5.5 Hz, 1H), 8.34 (s, 1H); MS: (M + H) ⁺ 198.
Process 3 :
Modification : Using 208 mg of 1,7-dichloro-isoquinoline, 350 mg of product was obtained (89% (yield)).
Product :

データ：MS：(M+Na)⁺ 415.

Data: MS: (M + Na) ⁺ 415.

  The following intermediates can be prepared as described herein and incorporated into compounds of Formula 1.

工程1：
修飾：25gの4-メチル桂皮酸を用い、15.3gの生成物を得た(62%(収率))。
生成物：

Process 1 :
Modification : Using 25 g of 4-methylcinnamic acid, 15.3 g of product was obtained (62% (yield)).
Product :

データ：¹H NMR (400MHz、CD₃OD) δ ppm 2.50 (s、3H)、6.54 (d、J=7.1Hz、1H)、7.13 (d、J=7.1Hz、1H)、7.49 (m、2H)、8.22 (s、1H)、11.49 (s、1H)；MS：(M+H)⁺ 160.
工程2：
修飾：15.3gの7-メチル-2H-イソキノリン-1-オンを用い、5.15gの生成物を得た(30%(収率))。
生成物：

Data: ¹ H NMR (400MHz, CD ₃ OD) δ ppm 2.50 (s, 3H), 6.54 (d, J = 7.1Hz, 1H), 7.13 (d, J = 7.1Hz, 1H), 7.49 (m, 2H) ), 8.22 (s, 1H), 11.49 (s, 1H); MS: (M + H) ⁺ 160.
Process 2 :
Modification : Using 15.3 g of 7-methyl-2H-isoquinolin-1-one, 5.15 g of product was obtained (30% (yield)).
Product :

データ：¹H NMR (400MHz、CDCl₃) δ ppm 2.58 (s、3H)、7.56 (m、2H)、7.73 (d、J=8.3Hz、1H)、8.09 (s、1H)、8.20 (d、J=5.6Hz、1H)；MS：(M+H)⁺ 178.
工程3：
修飾：205mgの1-クロロ-7-メチル-イソキノリンを用い、350mgの生成物を得た(89 %(収率))。
生成物：

Data: ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 2.58 (s, 3H), 7.56 (m, 2H), 7.73 (d, J = 8.3 Hz, 1H), 8.09 (s, 1H), 8.20 (d, J = 5.6Hz, 1H); MS: (M + H) ⁺ 178.
Process 3 :
Modification : Using 205 mg of 1-chloro-7-methyl-isoquinoline, 350 mg of product was obtained (89% (yield)).
Product :

データ：MS：(M+H)⁺ 373.

Data: MS: (M + H) ⁺ 373.

  The following intermediates can be prepared as described herein and incorporated into compounds of Formula 1.

工程1：
修飾： 33gの4-メトキシ桂皮酸を用い、7gの生成物を得た(33%(収率))。
生成物：

Process 1 :
Modification : Using 33 g of 4-methoxycinnamic acid, 7 g of product was obtained (33% (yield)).
Product :

データ：¹H NMR (500MHz、CD₃COCD₃) δ ppm 3.90 (s、3H)、6.49 (d、J=7.0Hz、1H)、7.10 (d、J=7.3Hz、1H)、7.28 (dd、J=8.6、2.8Hz、1H)、7.57 (d、J=8.9Hz、1H)、7.71 (d、J=2.8Hz、1H).
工程2：
修飾： 4gの7-メトキシ-2H-イソキノリン-1-オンを用い、3gの生成物を得た(68 %(収率))。
生成物：

Data: ¹ H NMR (500 MHz, CD ₃ COCD ₃ ) δ ppm 3.90 (s, 3H), 6.49 (d, J = 7.0 Hz, 1H), 7.10 (d, J = 7.3 Hz, 1H), 7.28 (dd, J = 8.6, 2.8Hz, 1H), 7.57 (d, J = 8.9Hz, 1H), 7.71 (d, J = 2.8Hz, 1H).
Process 2 :
Modification : 4 g of 7-methoxy-2H-isoquinolin-1-one was used to give 3 g of product (68% (yield)).
Product :

データ：¹H NMR (400MHz、CDCl₃) δ ppm 3.98 (s、3H)、7.38 (dd、J=8.9、2.6Hz、1H)、7.52 (m、2H)、7.73 (d、J=8.8Hz、1H)、8.16 (d、J=5.4Hz、1H).
工程3：
修飾： 533mgの1-クロロ-7-メトキシ-イソキノリンを用い、1115mgの生成物を得た(100 %(収率))。
生成物：

Data: ¹ H NMR (400MHz, CDCl ₃ ) δ ppm 3.98 (s, 3H), 7.38 (dd, J = 8.9, 2.6Hz, 1H), 7.52 (m, 2H), 7.73 (d, J = 8.8Hz, 1H), 8.16 (d, J = 5.4Hz, 1H).
Process 3 :
Modification : 533 mg of 1-chloro-7-methoxy-isoquinoline was used to give 1115 mg of product (100% (yield)).
Product :

  The following intermediates can be prepared as described herein and incorporated into compounds of Formula 1.

工程1：
修飾： 19.6gの4-フルオロ-3-メトキシ桂皮酸を用い、9.5gの生成物を得た(48%(収率))。
生成物：

Process 1 :
Modification : Using 19.6 g of 4-fluoro-3-methoxycinnamic acid, 9.5 g of product was obtained (48% (yield)).
Product :

データ：¹H NMR (400MHz、CD₃COCD₃) δ ppm 4.00 (s、1H)、6.49 (d、J=7.34Hz、1H)、7.19 (d、J=7.09Hz、1H)、7.29 (d、J=8.07Hz、1H)、7.86 (d、J=11.74Hz、1H).
工程2：
修飾： 9gの7-フルオロ-6-メトキシ-2H-イソキノリン-1-オンを用い、7gの生成物を得た(70%(収率))。
生成物：

Data: ¹ H NMR (400 MHz, CD ₃ COCD ₃ ) δ ppm 4.00 (s, 1H), 6.49 (d, J = 7.34Hz, 1H), 7.19 (d, J = 7.09Hz, 1H), 7.29 (d, J = 8.07Hz, 1H), 7.86 (d, J = 11.74Hz, 1H).
Process 2 :
Modification : Using 9 g of 7-fluoro-6-methoxy-2H-isoquinolin-1-one, 7 g of product was obtained (70% yield).
Product :

データ：¹H NMR (400MHz、CDCl₃) δ ppm 4.04 (s、3H)、7.17 (d、J=8.07Hz、1H)、7.48 (d、J=5.62Hz、1H)、7.94 (d、J=11.49Hz、1H)、8.20 (d、J=5.62Hz、1H).
工程3：
修飾： 222mgの1-クロロ-7-フルオロ-6-メトキシ-イソキノリンを用い、406mgの生成物を得た。
生成物：

Data: ¹ H NMR (400MHz, CDCl ₃ ) δ ppm 4.04 (s, 3H), 7.17 (d, J = 8.07Hz, 1H), 7.48 (d, J = 5.62Hz, 1H), 7.94 (d, J = 11.49Hz, 1H), 8.20 (d, J = 5.62Hz, 1H).
Process 3 :
Modification: 222 mg of 1-chloro-7-fluoro-6-methoxy-isoquinoline was used to give 406 mg of product.
Product :

  The following intermediates can be prepared as described herein and incorporated into compounds of Formula 1.

工程1：
修飾： 3.8gの3-(2,3-ジヒドロ-ベンゾフラン-7-イル)-アクリル酸を用い、2gの生成物を得た(53%(収率))。
生成物：

Process 1 :
Modification : 3.8 g of 3- (2,3-dihydro-benzofuran-7-yl) -acrylic acid was used to give 2 g of product (53% (yield)).
Product :

データ： ¹H NMR (400MHz、CD₃OD) δ ppm 3.37 (t、J=9.05Hz、1H)、4.73 (t、J=9.05Hz、2H)、6.67 (d、J=7.09Hz、1H)、7.10 (d、J=7.09Hz、1H)、7.37 (d、J=8.07Hz、1H)、7.81 (d、J=8.07Hz、1H)；MS：(M+H)⁺ 188.
工程2：
修飾： 1.87gの2,3-ジヒドロ-7H-フロ[2,3-f]イソキノリン-6-オンを用い、1.84gの生成物を得た(90%(収率))。
生成物：

Data: ¹ H NMR (400 MHz, CD ₃ OD) δ ppm 3.37 (t, J = 9.05Hz, 1H), 4.73 (t, J = 9.05Hz, 2H), 6.67 (d, J = 7.09Hz, 1H), 7.10 (d, J = 7.09Hz, 1H), 7.37 (d, J = 8.07Hz, 1H), 7.81 (d, J = 8.07Hz, 1H); MS: (M + H) ⁺ 188.
Process 2 :
Modification : 1.87 g of 2,3-dihydro-7H-furo [2,3-f] isoquinolin-6-one was used to give 1.84 g of product (90% (yield)).
Product :

データ：¹H NMR (400Hz、CDCl₃) δ ppm 3.43 (t、J=9.05Hz、2H)、4.82 (t、J=9.05Hz、2H)、7.52 (d、J=8.56Hz、1H)、7.66 (d、J=5.62Hz、1H)、7.84 (d、J=8.31Hz、1H)、8.19 (d、J=5.62Hz、1H)；MS (M+H)⁺ 206.
工程3：
修飾： 206mgの6-クロロ-2,3-ジヒドロ-フロ[2,3-f]イソキノリンを用い、300mgの生成物混合物を得た。
生成物：

Data: ¹ H NMR (400Hz, CDCl ₃ ) δ ppm 3.43 (t, J = 9.05Hz, 2H), 4.82 (t, J = 9.05Hz, 2H), 7.52 (d, J = 8.56Hz, 1H), 7.66 (d, J = 5.62Hz, 1H), 7.84 (d, J = 8.31Hz, 1H), 8.19 (d, J = 5.62Hz, 1H); MS (M + H) ⁺ 206.
Process 3 :
Modification : Using 206 mg 6-chloro-2,3-dihydro-furo [2,3-f] isoquinoline, 300 mg product mixture was obtained.
Product :

  The following intermediates can be prepared as described herein and incorporated into compounds of Formula 1.

工程1：
修飾： 1.14gの3-(2,3-ジヒドロ-ベンゾフラン-4-イル)-アクリル酸を用い、600mgの生成物を得た(52%(収率))。
生成物：

Process 1 :
Modification : 1.14 g of 3- (2,3-dihydro-benzofuran-4-yl) -acrylic acid was used to give 600 mg of product (52% yield).
Product :

データ：¹H NMR (400MHz、CD₃OD) δ ppm 3.35 (t、J=8.93Hz、2H)、4.74 (t、J=8.93Hz、2H)、6.49 (d、J=7.09Hz、1H)、6.95 (d、J=8.56Hz、1H)、7.25 (d、J=7.09Hz,、1H)、8.13 (d、J=8.80Hz、1H)；MS (M+H)⁺ 188.
工程2：
修飾： 560mgの1,7-ジヒドロ-2H-フロ[3,2-f]イソキノリン-6-オンを用い、380mgの生成物を得た(48%(収率))。
生成物：

Data: ¹ H NMR (400 MHz, CD ₃ OD) δ ppm 3.35 (t, J = 8.93 Hz, 2H), 4.74 (t, J = 8.93 Hz, 2H), 6.49 (d, J = 7.09 Hz, 1H), 6.95 (d, J = 8.56Hz, 1H), 7.25 (d, J = 7.09Hz, 1H), 8.13 (d, J = 8.80Hz, 1H); MS (M + H) ⁺ 188.
Process 2 :
Modification : 560 mg of 1,7-dihydro-2H-furo [3,2-f] isoquinolin-6-one was used to give 380 mg of product (48% (yield)).
Product :

データ：¹H NMR (400Hz、CDCl₃) δ ppm 3.47 (t、J=9.05Hz、2H)、4.84 (t、J=9.05Hz、2H)、7.24 (d、J=8.56Hz、1H)、7.33 (d、J=5.87Hz、1H)、8.20 (m、2 H)；MS (M+H)⁺ 206.
工程3：
修飾：105mgの6-クロロ-1,2-ジヒドロ-フロ[3,2-f]イソキノリンを用い、390mgの生成物混合物を得た。
生成物：

Data: ¹ H NMR (400Hz, CDCl ₃ ) δ ppm 3.47 (t, J = 9.05Hz, 2H), 4.84 (t, J = 9.05Hz, 2H), 7.24 (d, J = 8.56Hz, 1H), 7.33 (d, J = 5.87Hz, 1H), 8.20 (m, 2H); MS (M + H) ⁺ 206.
Process 3 :
Modification : 105 mg of 6-chloro-1,2-dihydro-furo [3,2-f] isoquinoline was used to give 390 mg of product mixture.
Product :

  The following intermediates can be prepared as described herein and incorporated into compounds of Formula 1.

工程1：
修飾：MeCN (10mL)中の6-メトキシ-2H-イソキノリン-1-オン(700mg)およびNCS (532mg)の化合物を3時間還流した。ろ過して600mg(72%)の所望の生成物を固体として得た。
生成物：

Process 1 :
Modification : A compound of 6-methoxy-2H-isoquinolin-1-one (700 mg) and NCS (532 mg) in MeCN (10 mL) was refluxed for 3 hours. Filtration gave 600 mg (72%) of the desired product as a solid.
Product :

データ：¹H NMR(400MHz、CD₃OD) δ ppm 3.96 (s、1H)、7.19 (dd、J=8.80、2.45Hz、1H)、7.28 (d、J=2.45Hz、1H)、7.34 (s、1H)、8.25 (d、J=9.05Hz、1H)；MS：(M+H)⁺ 210.
工程2：
修飾：500mgの4-クロロ-6-メトキシ-2H-イソキノリン-1-オンを用い、400mgの生成物を得た。
生成物：

Data: ¹ H NMR (400 MHz, CD ₃ OD) δ ppm 3.96 (s, 1H), 7.19 (dd, J = 8.80, 2.45 Hz, 1H), 7.28 (d, J = 2.45 Hz, 1H), 7.34 (s , 1H), 8.25 (d, J = 9.05Hz, 1H); MS: (M + H) + 210.
Process 2 :
Modification : 400 mg of product was obtained using 500 mg of 4-chloro-6-methoxy-2H-isoquinolin-1-one.
Product :

データ：¹H NMR (400Hz、CDCl₃) δ ppm 4.01 (s、3H)、7.35 (d、J=2.45Hz、1H)、7.41 (d、J=2.45Hz、1H)、8.24 (d、J=9.29Hz、1H)、8.27 (s、1H)；MS：(M+H)⁺ 229.
方法B

Data: ¹ H NMR (400Hz, CDCl ₃ ) δ ppm 4.01 (s, 3H), 7.35 (d, J = 2.45Hz, 1H), 7.41 (d, J = 2.45Hz, 1H), 8.24 (d, J = 9.29Hz, 1H), 8.27 (s, 1H); MS: (M + H) ⁺ 229.
Method B

工程1:

Process 1 :

  A mixture of 4-methoxy-2-methyl-benzoic acid (5.00 g, 30.1 mmol) and thionyl chloride (20.0 g, 0.17 mol) was heated to reflux for 30 minutes. Volatiles were removed under reduced pressure. After evacuating overnight, the viscous oily acid chloride was used in the next reaction as a crude material without further purification.

To a solution of 4-methoxy-2-methyl-benzyl chloride in CH ₂ Cl ₂ (60 mL) at 0 ° C., diethylamine was added dropwise. The formed mixture was allowed to warm to ambient temperature with stirring for 2 hours. Volatiles were removed under reduced pressure. The residue was triturated with EtOAc (100 mL) and then filtered. The filtrate was washed with 1M HCl, 1M NaOH, and brine and dried over MgSO ₄ . The solvent was evaporated to give 6.51 g (98%) of the desired product as a viscous oil.
LC-MS (retention time: 1.20 min, method B), MS m / z 222 (M + + H).
Process 2 :

To a solution of N, N-diethyl-4-methoxy-2-methyl-benzamide (221 mg, 1.0 mmol) in THF (2 mL) at −78 ° C. was added n-BuLi (0.84 mL, 2.5 M in hexane, 2.10 mmol). added. The orange solution formed was kept at this temperature for a further 30 minutes and then benzonitrile (103 mg, 1.0 mmol) was added dropwise. The final solution was allowed to warm to ambient temperature overnight with stirring. Attenuated with ice-cold 5% citric acid. Filtered, washed with water and then dried. Trituration with 2: 1 hexane-EtOAc (5 mL) gave 205 mg (82%) of the desired product as a white solid.
¹ H NMR (d ₆ -DMSO) δ 3.89 (s, 3H), 6.84 (s, 1H), 7.05-7.07 (m, 1H), 7.18 (d, J = 2.5Hz, 1H), 7.44-7.51 (m 3H), 7.78 (d, J = 7.0Hz, 1H), 8.11 (d, J = 9.0Hz, 1H);
LC-MS (retention time: 1.20 min, method B), MS m / z 252 (M + + H).
Process 3 :

This product, 1-chloro-6-methoxy-3-phenyl-isoquinoline, was prepared in the same manner as above except that 6-methoxy-3-phenyl-2H-isoquinolin-1-one was used instead.
¹ H NMR (CDCl ₃ ) δ 3.97 (s, 3H), 7.12 (d, J = 2.5Hz, 1H), 7.23-7.26 (m, 1H), 7.40-7.42 (m, 1H), 7.46-7.50 (m , 2H), 7.89 (s, 1H), 8.08 (d, J = 7.0Hz, 2H), 8.21 (d, J = 9.0Hz, 1H); LC-MS (retention time: 1.90min, method B), MS ^{m / z 270,271 (M + +} H).

  The following intermediates can be prepared as described herein and incorporated into compounds of Formula 1.

To a solution of N, N-diethyl-4-methoxy-2-methyl-benzamide (332 mg, 1.5 mmol) in THF (15 mL) at −78 ° C. was added t-BuLi (1.7 M solution in pentane, 1.3 mL, 2.25 mmol). ) Was added. The resulting red solution was stirred at −78 ° C. for 10 min and then 2-cyanopyridine (156 mg, 1.5 mmol) was added. The reaction mixture was then warmed to room temperature and stirred overnight. The reaction was quenched with saturated NH ₄ Cl solution and then extracted twice with ethyl acetate. The combined organic layer was dried (MgSO ₄ ) and then concentrated. The crude product was purified by Prep. HPLC to give a yellowish solid as a TFA salt (85 mg, 15% (yield)).
¹ H NMR (400 MHz, CD ₃ OD) δ 3.91 (m, 3H), 7.09 (dd, J = 9.05, 2.45 Hz, 1H), 7.17 (d, J = 2.45 Hz, 1H), 7.37 (s, 1H) , 7.42 (m, 1H), 7.92 (m, 1H), 8.08 (d, J = 8.07Hz, 1H), 8.18 (d, J = 9.05Hz, 1H), 8.65 (d, J = 4.89Hz, 1H) .
LC-MS (retention time: 2.14 min.), MS m / z 253 (MH +).
Process 2 (Scheme 3, Process 1):

6-Methoxy-3-pyridin-2-yl-2H-isoquinolin-1-one TFA salt (85 mg, 0.232 mmol) was heated to reflux with POCl ₃ (3.0 mL) for 2 days. Then POCl ₃ was distilled off and the residue was quenched with ice. It was then neutralized with 10N NaOH solution and the brown solid was recovered as a pure product (62 mg, 99% (yield)).
LC-MS (retention time: 2.063min.), MS m / z 271 (MH +).

  The following intermediates can be prepared as described herein and incorporated into compounds of Formula 1.

To a solution of N, N-diethyl-4-methoxy-2-methyl-benzamide (332 mg, 1.5 mmol) in THF (15 mL) at −78 ° C. was added t-BuLi (1.7 M solution in pentane, 1.3 mL, 2.25 mmol). ) Was added. The resulting red solution was stirred at −78 ° C. for 10 minutes and then 4-cyanopyridine (164 mg, 1.575 mmol) was added. The reaction mixture was then warmed to room temperature and stirred overnight. The reaction was quenched with saturated NH ₄ Cl solution and the yellow precipitate was collected as a pure product (145 mg, 38% (yield)).
¹ H NMR (CD ₃ OD, 400 MHz) δ 3.91 (s, 3H), 7.18 (dd, J = 8.8 Hz, 2.8 Hz, 1H), 7.26 (m, 2H), 8.06 (d, J = 6.0 Hz, 2H ), 8.16 (d, J = 8.8Hz, 1H), 8.84 (d, J = 6.0Hz, 2H).
LC-MS (retention time: 1.300 min.), MS m / z 253 (MH +).
Process 2 (Scheme 3, Process 1):

6-Methoxy-3-pyridin-4-yl-2H-isoquinolin-1-one (134 mg, 0.531 mmol) was heated to reflux with POCl ₃ (6.0 mL) for 5 days. Then POCl ₃ was distilled off and the residue was then quenched with ice water. It was then neutralized with saturated NaHCO ₃ solution and the brown solid was recovered as a pure product (125 mg, 87% (yield)).
¹ H NMR (DMSO-d6, 400 MHz) δ 3.99 (s, 3H), 7.53 (dd, J = 9.04 Hz, 2.44 Hz, 1H), 7.59 (d, J = 2.69 Hz, 1H), 8.26 (d, J = 9.05Hz, 1H), 8.30 (d, J = 5.38Hz, 2H), 8.73 (s, 1H), 8.85 (d, J = 6.36Hz, 2H).
LC-MS (retention time: 2.027 min.), MS m / z 271 (MH +).

  The following intermediates can be prepared as described herein and incorporated into compounds of Formula 1.

To a solution of N, N-diethyl-4-methoxy-2-methyl-benzamide (332 mg, 1.5 mmol) in THF (15 mL) at −78 ° C. was added t-BuLi (1.7 M solution in pentane, 1.3 mL, 2.25 mmol). ) Was added. The resulting red solution was stirred at −78 ° C. for 10 minutes and then 4-dimethylaminobenzonitrile (219 mg, 1.5 mmol) was added. The reaction mixture was then warmed to room temperature and stirred overnight. The reaction was quenched with saturated NH ₄ Cl solution and the yellow precipitate was collected and then triturated with ether to give an off-white solid as a pure product (247 mg, 56% (yield)).
¹ H NMR (DMSO-d6, 400 MHz) δ 2.97 (s, 6H), 3.87 (s, 3H), 6.72 (s, 1H), 6.78 (d, J = 8.80Hz, 2H), 6.97 (dd, J = 8.80, 2.45Hz, 1H), 7.10 (d, J = 2.45Hz, 1H), 7.65 (d, J = 8.80Hz, 2H), 8.05 (d, J = 8.80Hz, 1H), 11.11 (s, 1H) .
LC-MS (retention time: 2.023 min.), MS m / z 295 (MH +).

3- (4-Dimethylamino-phenyl) -6-methoxy-2H-isoquinolin-1-one (245 mg, 0.83 mmol) was heated to reflux with POCl ₃ (10.0 mL) for 2 days. Then POCl ₃ was distilled off and the residue was quenched with ice. It was then neutralized with 10 N NaOH solution and then extracted twice with ethyl acetate. The organic layers were mixed and then dried (MgSO ₄ ). The solvent was evaporated to give an orange solid as product (215 mg, 83% (yield)).
¹ H NMR (400 MHz, CD ₃ OD) δ 3.01 (s, 6H), 3.96 (s, 3H), 6.88 (d, J = 9.05Hz, 2H), 7.20 (dd, J = 9.17, 2.57Hz, 1H) 7.28 (d, J = 2.45Hz, 1H), 7.94 (s, 1H), 7.96 (d, J = 9.05Hz, 2H), 8.13 (d, J = 9.29Hz, 1H).
LC-MS (retention time: 2.543min.), MS m / z 313 (MH +).

A mixture of [4- (1-chloro-6-methoxy-isoquinolin-3-yl) -phenyl] -dimethyl-amine (110 mg, 0.35 mmol) and tetrabutylphosphonium hydrogen difluoride (0.5 g) in a Smith electromagnetic reactor Heated to 140 ° C. for minutes. Next, water was added thereto and extracted with ethyl acetate. The organic layer was separated, washed with water and then dried (MgSO ₄ ). The solvent was evaporated to give a brownish solid as product (85 mg, 82% (yield)).
LC-MS (retention time: 2.320 min.), MS m / z 297 (MH +).
Method C

工程1:

Process 1 :

To a solution of N-BOC-3- (R) -hydroxy-L-proline (6.22 g, 26.9 mmol) in DMF (250 mL) at 0 ° C., NaH (60%, 3.23 g, 80.8 mmol) was added several times. Added separately. The formed suspension was stirred at this temperature for 30 minutes. 1,3-Dichloro-isoquinoline (5.33 g, 26.9 mmol) was added in one portion as a solid and the final mixture was stirred for 12 hours at ambient temperature. Attenuated with ice-cold 5% citric acid (aq) and extracted with EtOAC (300 mL). The aqueous phase was extracted again with EtOAC. The combined organic layers were each washed with 5% citric acid (aq) and brine, dried over MgSO ₄ and then filtered. The filtrate was evaporated to dryness under reduced pressure and 10.53 g (99.8%) of 4- (6-methoxy-isoquinolin-1-yloxy) -pyrrolidine-1,2-dicarboxylic acid 1-tert-butyl ester off-white Obtained as a foam. This material was used in the next step reaction as a crude material without further purification.
¹ H NMR (CD ₃ OD) δ 1.43, 1.44 (rotamers, 9H), 2.39-2.44 (m, 1H), 2.68-2.72 (m, 1H), 3.80-3.90 (m, 2H), 4.44-4.52 (m, 1H), 5.77 (b, 1H), 7.39 (s, 1H), 7.58 (t, J = 7.3Hz, 1H), 7.71-7.78 (m, 2H), 8.16 (d, J = 7.5Hz, 1H);
LC-MS (retention time: 1.80min, method B), MS m / z 392 (M + + H).
Process 2 :

4- (6-Methoxy-isoquinolin-1-yloxy) -pyrrolidine-1,2-dicarboxylic acid 1-tert-butyl ester (39 mg, 0.10 mmol), phenylboronic acid (14.6 mg, 0.12) in THF (2 mL) mmol), sodium tert-butoxide (38 mg, 0.40 mmol), and ((t-Bu) ₂ POH) ₂ PdCl ₂ (POPd) (5 mg, 0.01 mmol) were heated to reflux for 4 hours. After cooling, the formed mixture was quenched with 5% citric acid (aq) and then extracted with EtOAc (20 mL). The organic layer was washed with brine, dried over MgSO ₄ , filtered and evaporated. The residue was purified by prep-HPLC to give 36 mg (83%) of the desired product as an off-white foam.
¹ H NMR (CD ₃ OD) δ 1.43, 1.45 (rotamers, 9H), 2.51-2.56 (m, 1H), 2.74-2.82 (m, 1H), 3.88-3.92 (m, 1H), 3.98-4.01 (m, 1H), 4.50-4.57 (m, 1H), 5.95 (b, 1H), 7.36-7.39 (m, 1H), 7.45-7.48 (m, 2H), 7.55 (t, J = 7.3Hz, 1H ), 7.70 (t, J = 7.5Hz, 1H), 7.84-7.89 (m, 2H), 8.14-8.17 (m, 3H), 9.05 (b, 1H);
LC-MS (retention time: 1.97min, method B), MS m / z 435 (M + + H).

  The following intermediates can be prepared as described herein and incorporated into compounds of Formula 1.

4-メトキシフェニルボロン酸を用いて製造した。
¹H NMR (CD₃OD) δ 1.40、1.45 (回転異性体、9H)、2.50-2.55 (m、1H)、2.73-2.81 (m、1H)、3.81-3.89 (m、4H)、3.98-4.01 (m、1H)、4.50-4.57 (m、1H)、5.93 (b、1H)、7.02 (d、J=9.0Hz、2H)、7.50 (t、J=7.3Hz、1H)、7.67 (t、J=7.5Hz、1H)、7.73 (s、1H)、7.83 (d、J=8.5Hz、1H)、8.09 (d、J=8.5Hz、2H)、8.15 (d、J=8.0Hz、1H);
LC-MS (保持時間：2.00min、方法B)、MS m/z 465 (M⁺+H).

Prepared using 4-methoxyphenylboronic acid.
¹ H NMR (CD ₃ OD) δ 1.40, 1.45 (rotamer, 9H), 2.50-2.55 (m, 1H), 2.73-2.81 (m, 1H), 3.81-3.89 (m, 4H), 3.98-4.01 (m, 1H), 4.50-4.57 (m, 1H), 5.93 (b, 1H), 7.02 (d, J = 9.0Hz, 2H), 7.50 (t, J = 7.3Hz, 1H), 7.67 (t, J = 7.5Hz, 1H), 7.73 (s, 1H), 7.83 (d, J = 8.5Hz, 1H), 8.09 (d, J = 8.5Hz, 2H), 8.15 (d, J = 8.0Hz, 1H) ;
LC-MS (retention time: 2.00 min, method B), MS m / z 465 (M + + H).

  The following intermediates were prepared as described above.

Prepared using 4-pyridylboronic acid.
¹ H NMR (CD ₃ OD) δ 1.43, 1.46 (rotamer, 9H), 2.53-2.56 (m, 1H), 2.80-2.89 (m, 1H), 3.90-3.93 (m, 1H), 4.00-4.05 (m, 1H), 4.50-4.57 (m, 1H), 6.00, 6.05 (rotamers, 1H), 7.80 (t, J = 7.3Hz, 1H), 7.87 (t, J = 7.5Hz, 1H), 8.08 (d, J = 8.5Hz, 1H), 8.32 (d, J = 8.0Hz, 1H), 8.49 (s, 1H), 8.84 (d, J = 6.0Hz, 2H), 8.84 (d, J = 6.5 Hz, 2H);
LC-MS (retention time: 1.39min, method B), MS m / z 436 (M + + H).

  The following intermediates can be prepared as described herein and incorporated into compounds of Formula 1.

4-N,N-ジメチルアミノ-フェニルボロン酸を用いて製造した。
LC-MS (保持時間：1.64min、方法B)、MS m/z 478 (M⁺+H).
方法D

Prepared using 4-N, N-dimethylamino-phenylboronic acid.
LC-MS (retention time: 1.64min, method B), MS m / z 478 (M + + H).
Method D

工程1:

Process 1 :

To a solution of N, N-diethyl-4-methoxy-2-methyl-benzamide (633 mg, 2.9 mmol) in THF (15 mL) at −78 ° C. was added n-BuLi (2.3 mL, 2.5 M in hexane, 5.74 mmol). Added dropwise. The formed red solution was maintained at this temperature for an additional 30 minutes and then thiazole-2-carboxylic acid ethyl ester (A. Medici et al, Tetrahedron Lett. 1983, p2901) in THF (5 mL) at −78 ° C. ( 450 mg, 2.9 mmol) solution was cannulated. The final dark green solution was maintained at this temperature for 2 hours with stirring. Attenuated with saturated NH ₄ Cl (aq) and then extracted with EtOAc (50 mL). The organic layer was washed with saturated NH ₄ Cl (aq) and brine, dried and purified by flash column chromatography eluting with 2: 1 EtOAc: hexanes to yield 405 mg (45%) of the desired product as an off-white viscous oil. Obtained as a thing.
¹ H NMR (CDCl ₃ ) δ 1.08 (t, J = 7.0Hz, 6H), 3.22 (b, 2H), 3.44 (b, 2H), 3.79 (s, 3H), 4.59 (s, 2H), 6.79- 6.81 (m, 1H), 6.86 (d, J = 2.5Hz, 1H), 7.16 (d, J = 8.5Hz, 1H), 7.66 (d, J = 3.0Hz, 1H), 8.00 (d, J = 3.0 Hz, 1H);
LC-MS (retention time: 1.30min, method B), MS m / z 333 (M + + H).
Process 2 :

Seal a mixture of N, N-diethyl-4-methoxy-2- (2-oxo-2-thiazol-2-yl-ethyl) -benzamide (405 mg, 1.22 mmol) and NH ₄ OAc (3.0 g, 38.9 mmol) Heat to 140 ° C. for 1 hour in a tube. The molten solution was poured into ice cold water, filtered and the cake was washed thoroughly with water. The dry brown solid (240 mg, 76%) was used crude in the next reaction without further purification.
LC-MS (retention time: 1.24 min, method B), MS m / z 259 (M + + H).
Process 3 :

This product, 1-chloro-6-methoxy-3-thiazol-2-yl-isoquinoline, was used as described above except that 6-methoxy-3-thiazol-2-yl-2H-isoquinolin-1-one was used instead. It was manufactured as follows.
¹ H NMR (CDCl ₃ ) δ 3.97 (s, 3H), 7.16 (d, J = 4.0Hz, 1H), 7.27-7.31 (m, 1H), 7.46 (d, J = 5.0Hz, 1H), 7.93 ( d, J = 5.5Hz, 1H), 8.22 (d, J = 15.5Hz, 1H), 8.39 (s, 1H);
LC-MS (retention time: 1.66min, method B), MS m / z 277 (M + + H).
Process 4 :

This product was prepared in the same manner as above except that 1-chloro-6-methoxy-3-thiazol-2-yl-isoquinoline was used instead.
¹ H NMR (CD ₃ OD) δ 0.97-1.09 (m, 12H), 1.24-1.29 (m, 10H), 1.44-1.46 (m, 1H), 1.87-1.90 (m, 1H), 2.20-2.26 (m , 1H), 2.30-2.36 (m. 1H), 2.65-2.71 (m, 1H), 2.93-2.96 (m, 1H), 3.96 (s, 3H), 4.12-4.27 (m, 2H), 4.38-4.52 (m, 2H), 5.12 (d, J = 10.5Hz, 1H), 5.29 (d, J = 17.5Hz, 1H), 5.69-5.74 (m, 1H), 5.99 (b, 1H), 7.14 (d, J = 9.0Hz, 1H), 7.33 (s, 1H), 7.66 (d, J = 3.5Hz, 1H), 7.93 (d, J = 3.0Hz, 1H), 8.05 (s, 1H), 8.11 (d, J = 9.0Hz, 1H), 9.14 (b, 1H);
LC-MS (retention time: 1.89min, method B), MS m / z 797 (M + + H).

  The following intermediates can be prepared as described herein and incorporated into compounds of Formula 1.

6-methoxy-3- (3-methoxy-isoxazol-5-yl) -2H-isoquinolin-1-one was converted to N, N-diethyl-4-methoxy-2- [2- (3-methoxy-isoxazol-5 Prepared using -yl) -2-oxo-ethyl] -benzamide.
¹ H NMR (DMSO-d ₆ ) δ 3.89 (s, 3H), 3.97 (s, 3H), 7.01 (s, 1H), 7.14-7.16 (m, 2H), 7.43 (s, 1H), 8.13 (d , J = 8.5Hz, 1H);
LC-MS (retention time: 1.31min, method B), MS m / z 273 (M + + H).

1-chloro-6-methoxy-3- (3-methoxy-isoxazol-5-yl) -isoquinoline is converted to 6-methoxy-3- (3-methoxy-isoxazol-5-yl) -2H-isoquinolin-1-one It was manufactured using.
¹ H NMR (CDCl ₃ ) δ 3.97 (s, 3H), 4.04 (s, 3H), 6.60 (s, 1H), 7.17 (d, J = 2.5Hz, 1H), 7.31-7.33 (m, 1H), 8.02 (s, 1H), 8.23 (d, J = 9.0Hz, 1H);
LC-MS (retention time: 1.73min, method B), MS m / z 291,293 (M + + H)

  The following intermediates can be prepared as described herein and incorporated into compounds of Formula 1.

N, N-diethyl-4-methoxy-2- [2- (5-methoxy-oxazol-2-yl) -2-oxo-ethyl] -benzamide and 5-methoxy-oxazole-2-carboxylic acid ethyl ester Manufactured.
LC-MS (retention time: 1.24 min, method B), MS m / z 347 (M + + H).

6-Methoxy-3- (5-methoxy-oxazol-2-yl) -2H-isoquinolin-1-one was converted to N, N-diethyl-4-methoxy-2- [2- (5-methoxy-oxazole-2 Prepared using -yl) -2-oxo-ethyl] -benzamide.
¹ H NMR (DMSO-d6) δ 3.94 (s, 3H), 4.01 (s, 3H), 6.34 (s, 1H), 6.99 (d, J = 2.0Hz, 1H), 7.12-7.14 (m, 1H) 7.25 (s, 1H), 8.32 (d, J = 9.0Hz, 1H);
LC-MS (retention time: 1.22 min, method B), MS m / z 274 (M + + H).

1-chloro-6-methoxy-3- (5-methoxy-oxazol-2-yl) -isoquinoline is converted to 6-methoxy-3- (5-methoxy-oxazol-2-yl) -2H-isoquinolin-1-one It was manufactured using.
¹ H NMR (CDCl ₃ ) δ 3.96 (s, 3H), 4.00 (s, 3H), 6.34 (s, 1H), 7.12 (d, J = 2.5Hz, 1H), 7.28-7.31 (m, 1H), 8.13 (s, 1H), 8.23 (d, J = 9.0Hz, 1H);
LC-MS (retention time: 1.58min, method B), MS m / z 291,293 (M + + H).

  The following intermediates can be prepared as described herein and incorporated into compounds of Formula 1.

To a solution of N, N-diethyl-4-methoxy-2-methyl-benzamide (332 mg, 1.5 mmol) in THF (15 mL) at −78 ° C. was added t-BuLi (1.7 M solution in pentane, 2.12 mL, 3.6 mmol). ) Was added. The resulting red solution was stirred at −78 ° C. for 10 minutes, then N-methylnicotinate (206 mg, 1.5 mmol) was added. The reaction mixture was stirred at −78 ° C. for 2 hours. The reaction was then quenched with saturated NH ₄ Cl solution and then extracted twice with ethyl acetate. The combined organic layer was dried (MgSO ₄ ) and then concentrated. The crude product was purified by Prep. HPLC to give a yellowish high viscosity oil as TFA salt (124 mg, 19% (yield)).
LC-MS (retention time: 1.740 min.), MS m / z 349 (M + Na ⁺ ).

N, N-diethyl-4-methoxy-2- (2-oxo-2-pyridin-3-yl-ethyl) -benzamide (120 mg, 0.272 mmol) was heated with ammonium acetate (1 g) for 3 hours. It was then cooled and water was added. Extract with ethyl acetate and then separate the organic layer. It was then dried (MgSO ₄ ) and concentrated to give a brownish solid product (65 mg, 95% (yield)).
¹ H NMR (400MHz, DMSO-d6) δ 3.89 (s, 3H), 6.93 (s, 1H), 7.10 (dd, J = 8.80, 2.45Hz, 1H), 7.19 (d, J = 2.45Hz, 1H) 7.52 (dd, J = 7.46, 4.77Hz, 1H), 8.15 (m, 2H), 8.64 (dd, J = 4.89, 1.47Hz, 1H), 8.96 (d, J = 1.71Hz, 1H), 11.51 ( s, 1H).
LC-MS (retention time: 1.377 min.), MS m / z 253 (MH +).

6-メトキシ-3-ピリジン-3-イル-2H-イソキノリン-1-オン(65mg、0.258mmol)を7日間POCl₃ (2.5mL)と加熱還流した。次に、POCl₃を留去し、残渣を氷水で減衰させた。次に、それを10 N NaOH溶液で中和し、次いで酢酸エチルで2回抽出した。混合有機層を乾燥し(MgSO₄)、次いで濃縮して黄色固体の生成物を得た(27mg、39%(収率))。LC-MS (保持時間：2.090min.)、MS m/z 271 (MH+).

6-Methoxy-3-pyridin-3-yl-2H-isoquinolin-1-one (65 mg, 0.258 mmol) was heated to reflux with POCl ₃ (2.5 mL) for 7 days. Next, POCl ₃ was distilled off and the residue was attenuated with ice water. It was then neutralized with 10 N NaOH solution and then extracted twice with ethyl acetate. The combined organic layers were dried (MgSO ₄ ) and then concentrated to give a yellow solid product (27 mg, 39% (yield)). LC-MS (retention time: 2.090 min.), MS m / z 271 (MH +).

  The following intermediates can be prepared as described herein and incorporated into compounds of Formula 1.

To a solution of N, N-diethyl-4-methoxy-2-methyl-benzamide (332 mg, 1.5 mmol) in THF (15 mL) at −78 ° C. was added t-BuLi (1.7 M solution in pentane, 2.2 mL, 3.75 mmol). ) Was added. The resulting red solution was stirred at −78 ° C. for 10 minutes, then N, N-dimethylanthranilic acid methyl ester (269 mg, 1.5 mmol) was added. The reaction mixture was stirred at −78 ° C. for 2 hours. The reaction was then quenched with saturated NH ₄ Cl solution and then extracted twice with ethyl acetate. The combined organic layer was dried (MgSO ₄ ) and then concentrated. The crude product was purified by Prep. HPLC to give a yellowish high viscosity oil as product (256 mg, 46% (yield)).
¹ H NMR (400 MHz, CD ₃ OD) δ 0.99-1.13 (m, 6H), 3.23-3.31 (m, 8H), 3.39 (m, 2H), 3.82 (s, 3H), 4.35 (s, 2H), 6.91 (dd, J = 8.44, 2.57Hz, 1H), 6.99 (d, J = 2.45Hz, 1H), 7.22 (d, J = 8.56Hz, 1H), 7.69 (t, J = 7.70Hz, 1H), 7.84 (m, 1H), 7.96 (d, J = 8.31Hz, 1H), 8.18 (d, J = 7.83Hz, 1H).
LC-MS (retention time: 1.557 min.), MS m / z 369 (MH +).

2- [2- (2-Dimethylamino-phenyl) -2-oxo-ethyl] -N, N-diethyl-4-methoxy-benzamide (250 mg, 0.678 mmol) was heated with ammonium acetate (1.5 g) for 2 hours. . It was then cooled and then water was added. Extraction with ethyl acetate separated the organic layer. It was then dried (MgSO ₄ ) and concentrated to give a yellowish solid as product (125 mg, 63% (yield)).
¹ H NMR (400 MHz, CD ₃ OD) δ 2.95 (s, 6H), 3.92 (s, 3H), 6.92 (s, 1H), 7.12 (dd, J = 8.80, 2.45Hz, 1H), 7.16 (d, J = 2.45Hz, 1H), 7.35 (m, 1H), 7.55 (m, 2H), 7.63 (d, J = 7.83Hz, 1H), 8.20 (d, J = 9.05Hz, 1H).
LC-MS (retention time: 2.097 min.), MS m / z 295 (MH +).

3- (2-Dimethylamino-phenyl) -6-methoxy-2H-isoquinolin-1-one (125 mg, 0.425 mmol) was heated to reflux with POCl ₃ (4.0 mL) for 1 day. Then POCl ₃ was distilled off and the residue was then quenched with ice. It was then neutralized with 10 N NaOH solution and then extracted twice with ethyl acetate. The organic layers were mixed and then dried (MgSO ₄ ). The solvent was evaporated to give a brownish solid as product (82 mg, 62% (yield)).
LC-MS (retention time: 2.040 min.), MS m / z 313 (MH ⁺ ).

A mixture of [2- (1-chloro-6-methoxy-isoquinolin-3-yl) -phenyl] -dimethyl-amine (82 mg, 0.262 mmol) and tetrabutylphosphonium hydrogen difluoride (1.0 g) was added in a Smith electromagnetic wave reactor. Heated to 140 ° C. for minutes. Then it was added water and then extracted with ethyl acetate. The organic layer was separated, washed with water and then dried (MgSO ₄ ). The solvent was evaporated to give the crude product, which was purified by Prep. HPLC to give a yellowish oily product (85 mg).
¹ H NMR (400 MHz, CD ₃ OD) δ 3.41 (s, 6H), 4.00 (s, 3H), 7.42 (dd, J = 9.05, 2.45 Hz, 1H), 7.53 (s, 1H), 7.71 (m, 2H), 7.99 (m, 1H), 8.16 (m, 2H), 8.31 (s, 1H).
LC-MS (retention time: 1.873 min.), MS m / z 297 (MH ⁺ ).

  The following intermediates can be prepared as described herein and incorporated into compounds of Formula 1.

To a solution of N, N-diethyl-4-methoxy-2-methyl-benzamide (332 mg, 1.5 mmol) in THF (15 mL) at −78 ° C. was added t-BuLi (1.7 M solution in pentane, 2.2 mL, 3.75 mmol) was added. The resulting red solution was stirred at −78 ° C. for 10 minutes and then (3-dimethylamino) benzoic acid methyl ester (269 mg, 1.5 mmol) was added. The reaction mixture was stirred at −78 ° C. for 2 hours. The reaction was then quenched with saturated NH ₄ Cl solution and then extracted twice with ethyl acetate. The combined organic layer was dried (MgSO ₄ ) and then concentrated. The crude product was purified by Prep. HPLC to give a yellowish high viscosity oil as TFA salt (245 mg, 33% (yield)).
¹ H NMR (400 MHz, CD ₃ OD) δ 1.01 (t, J = 6.85 Hz, 3H), 1.09 (m, 3H), 3.11 (s, 6H), 3.21 (m, 2H), 3.40 (m, 2H) , 3.79 (s, 3H), 4.39 (s, 2H), 6.84-6.91 (m, 2H), 7.19 (d, J = 8.32Hz, 1H), 7.35 (m, 1H), 7.49 (t, J = 8.07 Hz, 1H), 7.66-7.71 (m, 2 H).
LC-MS (retention time: 1.930 min.), MS m / z 369 (MH ⁺ ).

2- [2- (3-Dimethylamino-phenyl) -2-oxo-ethyl] -N, N-diethyl-4-methoxy-benzamide (240 mg, 0.497 mmol) was heated with ammonium acetate (2.0 g) for 2.5 hours. . It was then cooled and then water was added. It was recovered as a brownish solid pure product (95 mg, 65% (yield)).
¹ H NMR (400 MHz, CD ₃ OD) δ 2.98 (s, 6H), 3.88 (s, 3H), 6.74-6.87 (m, 2H), 7.01-7.07 (m, 3H), 7.18 (d, J = 2.44 Hz, 1H), 7.28 (t, J = 7.82Hz, 1H), 8.10 (d, J = 8.80Hz, 1H).
LC-MS (retention time: 1.773 min.), MS m / z 295 (MH ⁺ ).

3- (3-Dimethylamino-phenyl) -6-methoxy-2H-isoquinolin-1-one (92 mg, 0.312 mmol) was heated to reflux with POCl ₃ (3.0 mL) for 2 days. Then POCl ₃ was distilled off and the residue was then quenched with ice. It was then neutralized with saturated NaHCO ₃ solution and then extracted twice with ethyl acetate. The organic layers were mixed and then dried (MgSO ₄ ). The solvent was evaporated to give a brownish high viscosity oil as product (72 mg, 74% (yield)).
LC-MS (retention time: 2.297 min.), MS m / z 313 (MH ⁺ ).

A mixture of [3- (1-chloro-6-methoxy-isoquinolin-3-yl) -phenyl] -dimethylamine (72 mg, 0.23 mmol) and tetrabutylphosphonium hydrogen difluoride (0.5 g) was placed in a Smith electromagnetic wave reactor for 20 minutes. Heated to 140 ° C. Next, water was added thereto and extracted with ethyl acetate. The organic layer was separated, washed with water and then dried (MgSO ₄ ). The solvent was evaporated to give a brownish oil as product (58 mg, 85% (yield)).
LC-MS (retention time: 2.193 min.), MS m / z 297 (MH ⁺ ).

  The following intermediates can be prepared as described herein and incorporated into compounds of Formula 1.

エチルブロモピルベートを還流ジオキサン中エチルチオ尿素と縮合させてモノアルキルアミノチアゾールを定量的量のHBr塩として得た。2-エチルアミノ-チアゾール-4-カルボン酸エチルエステルをDMF中のEtIでアルキル化して2-ジエチルアミノ-チアゾール-4-カルボン酸エチルエステルを得た。LC/MS m/z 229 (MH)⁺
方法E

Ethyl bromopyruvate was condensed with ethyl thiourea in refluxing dioxane to give the monoalkylaminothiazole as a quantitative amount of HBr salt. 2-Ethylamino-thiazole-4-carboxylic acid ethyl ester was alkylated with EtI in DMF to give 2-diethylamino-thiazole-4-carboxylic acid ethyl ester. LC / MS m / z 229 (MH) ⁺
Method E

工程1:
モルホリン(5mL)中の2-シアノメチル-4-メトキシ-安息香酸メチルエステル(1.9gおよびTsOH. H₂O (0.15g、mmol)のサスペンジョンを4時間還流し、溶媒を減圧下で除去した。残渣をMeOH滴を用いてEtOAc/ヘキサンから再結晶し、生成物を得た(0.43g、17%)：LC-MS 保持時間：1.07 方法H)、MS m/z 266 (M⁺+1).
工程2:
POCl₃ (20mL)中の6-メトキシ-3-モルホリン-4-イル-イソキノリン-1-オール(0.298g、1.15mmol)の混合物を2時間還流し、溶媒を減圧下で除去し、次いで冷水を加えた。pHを1.0 N NaOHを加えて>11に調整した。水性層をEtOAcで抽出した。抽出物を乾燥し(MgSO₄)、溶媒を減圧下で除去して生成物を得た(0.299g、94%)：LC-MS 保持時間：1.68 方法H)、MS m/z 279 (M⁺+1).
工程3:
1-クロロ-6-メトキシ-3-モルホリン-4-イル-イソキノリン (0.050g、0.18mmol)およびテトラブチルホスホリウム水素ジフロリド(0.8g、2.8mmol)の混合物 [Synlett 1992、(4)、345-6]をマイクロ波で10分間140℃に加熱した。反応混合物をEtOAcで希釈し、上端にシリコンゲルの層を有するISCO 25gプレカラムでろ過し、次いで溶媒を除去して生成物を得た(0.037mg、77%)： ¹H NMR (クロロホルム-D) δ ppm 3.48 (m、4H)、3.84 (m、4H)、3.89 (s、3H)、6.46 (d、J=1.22Hz、1H)、6.85 (s、1H)、6.90 (dd、J=9.16、2.44Hz、1H)、7.82 (d、J=8.85Hz、1H). LC-MS 保持時間：1.56 方法H)、MS m/z 263 (M⁺+1).
方法F

Process 1 :
A suspension of 2-cyanomethyl-4-methoxy-benzoic acid methyl ester (1.9 g and TsOH.H ₂ O (0.15 g, mmol) in morpholine (5 mL) was refluxed for 4 hours and the solvent was removed under reduced pressure. Was recrystallized from EtOAc / hexanes using MeOH drops to give the product (0.43 g, 17%): LC-MS retention time: 1.07 Method H), MS m / z 266 (M ⁺ +1).
Process 2 :
A mixture of 6-methoxy-3-morpholin-4-yl-isoquinolin-1-ol (0.298 g, 1.15 mmol) in POCl ₃ (20 mL) was refluxed for 2 hours, the solvent was removed under reduced pressure, and then cold water was added. added. The pH was adjusted to> 11 by adding 1.0 N NaOH. The aqueous layer was extracted with EtOAc. The extract was dried (MgSO ₄ ) and the solvent removed under reduced pressure to give the product (0.299 g, 94%): LC-MS retention time: 1.68 Method H), MS m / z 279 (M ⁺ +1).
Process 3 :
A mixture of 1-chloro-6-methoxy-3-morpholin-4-yl-isoquinoline (0.050 g, 0.18 mmol) and tetrabutylphospholium hydrogen difluoride (0.8 g, 2.8 mmol) [Synlett 1992, (4), 345- 6] was heated to 140 ° C. in the microwave for 10 minutes. The reaction mixture was diluted with EtOAc and filtered through an ISCO 25 g precolumn with a layer of silicon gel on top, then the solvent was removed to give the product (0.037 mg, 77%): ¹ H NMR (chloroform-D) δ ppm 3.48 (m, 4H), 3.84 (m, 4H), 3.89 (s, 3H), 6.46 (d, J = 1.22Hz, 1H), 6.85 (s, 1H), 6.90 (dd, J = 9.16, 2.44Hz, 1H), 7.82 (d, J = 8.85Hz, 1H). LC-MS retention time: 1.56 Method H), MS m / z 263 (M ⁺ +1).
Method F

The 6-fluoro and 6-alkylisoquinolines used in the preparation of the compound of formula 1 were prepared via Pomeranz-Fritsch synthesis as follows (typical method: Preparation of optically active 8,8-disubstituted 1,1-biisoquinolin, K Hirao, R. Tsuchiya, Y. Yano, H. Tsue, Heterocycles 42 (1) 1996, 415-422). The product was converted to a 1-chloro derivative via an N-oxide intermediate.
General synthetic reaction formula

試薬および反応条件：(a)ベンゼン中で還流、水を共沸除去；(b)第一工程：エチルクロロホルメート、THF中亜リン酸トリメチル、第2工程：クロロホルム中チタニウムテトラクロリド；(c) CH₂Cl₂中MCPBA；(d) ベンゼン中POCl₃

6-イソプロポキシルおよび6-tert-ブトキシルイソキノリン中間体の製造:

Reagents and reaction conditions : (a) reflux in benzene, azeotropic removal of water; (b) first step: ethyl chloroformate, trimethyl phosphite in THF, second step: titanium tetrachloride in chloroform; (c ) MCPBA in CH ₂ Cl ₂ ; (d) POCl _{3 in} benzene

Preparation of 6-isopropoxyl and 6-tert-butoxylisoquinoline intermediates :

Direct ipso substitution of some 6-alkoxy-1-chloroisoquinolines with the corresponding alkoxide metal ions of 6-fluoro-1-chloroisoquinoline such as potassium tert-butoxide (53%) and sodium isopropoxide (54%) Manufactured by.
General synthetic reaction formula

R=アルコキシドアニオン、例えばtert-Bu、iso-Pr

R = alkoxide anion, eg tert-Bu, iso-Pr

Aromatic nucleophilic substitution of 6-fluoro-1-chloroisoquinoline with sodium isopropoxide and potassium tert-butoxide in DMF, each with the corresponding 6-isopropoxyl (54%): ¹ H as the main product NMR (400MHz, chloroform-d) δ ppm 1.43 (d, J = 6.11Hz, 6H) 4.76 (m, J = 6.11Hz, 1H) 7.08 (d, J = 2.45Hz, 1H) 7.29 (dd, J = 9.29, 2.45Hz, 1H) 7.50 (d, J = 5.62Hz, 1H) 8.18 (d, J = 5.87Hz, 1H) 8.24 (d, J = 9.29Hz, 1H), and 6-tert-butoxyl-1- Chloroisoquinoline (55%): ¹ H NMR (400 MHz, chloroform-d) δ ppm 1.48 (s, 9 H) 7.31 (m, 2 H) 7.47 (d, J = 5.62 Hz, 1H) 8.18 (d, J = 5.62 Hz, 1H) 8.21 (d, J = 9.78 Hz, 1H) was obtained. These were 6-alkoxyl-1-chloroisoquinolines incorporated into the compounds of formula 1 described herein.
Method G
General synthetic reaction formula

This synthesis method used techniques described in part in the following references:
(1) Hojo, Masaru; Masuda, Ryoichi; Sakaguchi, Syuhei; Takagawa, Makoto, Synthesis (1986), (12), 1016-17
(2) Rigby, James H .; Holsworth, Daniel D .; James, Kelly. Vinyl Isocyanates In Synthesis. [4 + 2] Cycloaddition Reactions With Benzyne Addends. Journal Of Organic Chemistry (1989), 54 (17), 4019- 20
(3) Uchibori, Y .; Umeno, M .; Yoshiokai, H .; Heterocycles, 1992, 34 (8), 1507-1510.
Step 1d : Preparation of racemic (1R, 2S) / (1S, 2R) -1-amino-2-vinylcyclopropanecarboxylic acid ethyl ester / hydrochloric acid (Method A and Method B) and N- (1R, 2S)- Chiral resolution of this racemate to produce 1-amino-2-vinylcyclopropanecarboxylic acid ethyl ester / hydrochloric acid (Method C)

The title compound was made a (step d) racemic by the following methods A and B respectively. The racemate was resolved to give chiral Boc- (1R, 2S) -1-amino-2-vinylcyclopropylcarboxylic acid ester, which was deprotected under acid conditions to give (1R, 2S) -1-amino- 2-Vinylcyclopropanecarboxylic acid ester / hydrochloric acid (Method C) was obtained.
Method A
A.1 Production of N-benzylimine of glycine ethyl ester

グリシンエチルエステル・塩酸 (303.8g、2.16mole)をtert-ブチルメチルエーテル(1.6L)に懸濁した。ベンズアルデヒド(231g、2.16mole)および無水硫酸ナトリウム(154.6g、1.09mole)を加え、次いで混合物を氷水浴を用いて0℃に冷却した。トリエチルアミン (455mL、3.26mole)を30分間かけて滴加し、次いで混合物を室温で48時間撹拌した。次に、反応物を氷冷水(1L)を加えて減衰させ、次いで有機層を分離した。水性相をtert-ブチルメチルエーテル(0.5L)で抽出し、混合有機相を飽和水性NaHCO₃ (1L)および塩水(1L)の混合物で洗浄した。溶液をMgSO₄で乾燥し、減圧下で濃縮して392.4gのN-ベンジルイミン生成物を高粘度黄色油状物として得、これを直接次の工程に用いた。¹H NMR (CDCl₃、300MHz) δ 1.32 (t、J=7.1Hz、3H)、4.24 (q、J=7.1Hz、2H)、4.41 (d、J=1.1Hz、2H)、7.39-7.47 (m、3H)、7.78-7.81 (m、2H)、8.31 (s、1H).
A.2 ラセミックN-Boc-(1R,2S)/(1S,2R)-1-アミノ-2-ビニルシクロプロパンカルボン酸エチルエステルの製造

Glycine ethyl ester / hydrochloric acid (303.8 g, 2.16 mole) was suspended in tert-butyl methyl ether (1.6 L). Benzaldehyde (231 g, 2.16 mole) and anhydrous sodium sulfate (154.6 g, 1.09 mole) were added and then the mixture was cooled to 0 ° C. using an ice water bath. Triethylamine (455 mL, 3.26 mole) was added dropwise over 30 minutes and then the mixture was stirred at room temperature for 48 hours. The reaction was then quenched by the addition of ice cold water (1 L) and then the organic layer was separated. The aqueous phase was extracted with tert-butyl methyl ether (0.5 L) and the combined organic phase was washed with a mixture of saturated aqueous NaHCO ₃ (1 L) and brine (1 L). The solution was dried over MgSO ₄ and concentrated under reduced pressure to give 392.4 g of N-benzylimine product as a highly viscous yellow oil which was used directly in the next step. ¹ H NMR (CDCl ₃ , 300 MHz) δ 1.32 (t, J = 7.1 Hz, 3H), 4.24 (q, J = 7.1 Hz, 2H), 4.41 (d, J = 1.1 Hz, 2H), 7.39-7.47 ( m, 3H), 7.78-7.81 (m, 2H), 8.31 (s, 1H).
A.2 Preparation of racemic N-Boc- (1R, 2S) / (1S, 2R) -1-amino-2-vinylcyclopropanecarboxylic acid ethyl ester

A suspension of lithium tert-butoxide (84.06 g, 1.05 mol) in dry toluene (1.2 L) was added to the glycine ethyl ester of N-benzylimine (100.4 g, 0.526 mol) and trans in dry toluene (0.6 L). A mixture of -1,4-bibromo-2-butene (107.0 g, 0.500 mol) was added dropwise over 60 minutes. After the addition was complete, the deep red mixture was damped by adding water (1 L) and tert-butyl methyl ether (TBME, 1 L). The aqueous phase was separated and extracted a second time with TBME (1 L). The organic layers were combined, 1N HCl (1 L) was added and then the mixture was stirred at room temperature for 2 hours. The organic layer was separated and then extracted with water (0.8 L). The aqueous phase was then mixed, saturated with salt (700 g), TBME (1 L) was added and the mixture was then cooled to 0 ° C. The stirred mixture was then basified to pH 14 by the dropwise addition of 10N NaOH, the organic layer was separated and the aqueous phase was then extracted with TBME (2 × 500 mL). The combined organic extracts were dried (MgSO ₄ ) and then concentrated to a volume of 1L. To this free amine solution was added BOC ₂ O or di-tert-butyl dicarbonate (131.0 g, 0.6 mol) and then the mixture was stirred at room temperature for 4 days. Further di-tert-butyl dicarbonate (50 g, 0.23 mol) was added to the reaction and the mixture was refluxed for 3 hours and then cooled to room temperature overnight. The reaction mixture was dried over MgSO ₄ and concentrated under reduced pressure to give 80 g of crude material. Racemic of 57g of a yellow oil which solidified on standing this residue in the refrigerator was purified by flash chromatography (2.5 elution with _{SiO 2, 1% ~2% MeOH} / CH 2 Cl 2 in Kg) (53%) N- Boc- (1R, 2S) / (1S, 2R) -1-Amino-2-vinylcyclopropanecarboxylic acid ethyl ester was obtained: ¹ H NMR (CDCl ₃ , 300 MHz) δ 1.26 (t, J = 7.1 Hz, 3H) , 1.46 (s, 9H), 1.43-1.49 (m, 1H), 1.76-1.82 (br m, 1H), 2.14 (q, J = 8.6Hz, 1H), 4.18 (q, J = 7.2Hz, 2H) , 5.12 (dd J = 10.3, 1.7Hz, 1H), 5.25 (br s, 1H), 5.29 (dd, J = 17.6, 1.7Hz, 1H), 5.77 (ddd, J = 17.6, 10.3, 8.9Hz, 1H ); MS m / z 254.16 (M-1).
A.3 Production of racemic (1R, 2S) / (1S, 2R) 1-amino-2-vinylcyclopropanecarboxylic acid ethyl ester / hydrochloric acid

N-Boc- (1R, 2S) / (1S, 2R) -1-amino-2-vinylcyclopropanecarboxylic acid ethyl ester (9.39 g, 36.8 mmol) was dissolved in 4N HCl / dioxane (90 ml, 360 mmol), Stir at room temperature for 2 hours. The reaction mixture was concentrated to obtain a quantitative amount of (1R, 2S) / (1S, 2R) -1-amino-2-vinylcyclopropanecarboxylic acid ethyl ester / hydrochloric acid (7 g, 100%). ¹ H NMR (methanol-d ₄ ) δ 1.32 (t, J = 7.1, 3H), 1.72 (dd, J = 10.2, 6.6 Hz, 1H), 1.81 (dd, J = 8.3, 6.6 Hz, 1H), 2.38 (q, J = 8.3Hz, 1H), 4.26-4.34 (m, 2H), 5.24 (dd, 10.3, 1.3Hz, 1H) 5.40 (d, J = 17.2, 1H), 5.69-5.81 (m, 1H) .
B.1 Production of racemic N-Boc-1-amino-2-vinylcyclopropanecarboxylic acid ethyl ester / hydrochloric acid
Method B

Commercial glycine ethyl ester of N, N-dibenzylimine (25.0 g, 93.53 mmol) in THF (112 mL) in a solution of potassium tert-butoxide (11.55 g, 102.9 mmol) in THF (450 mL) at -78 ° C. Was added. The reaction mixture was warmed to 0 ° C., stirred for 40 minutes and then cooled back to −78 ° C. To this solution, trans-1,4-dibromo-2-butene (20.0 g, 93.50 mmol) was added, and the mixture stirred at 0 ° C. for 1 hour was cooled to −78 ° C. Potassium tert-butoxide (11.55 g, 102.9 mmol) was added and the mixture was rapidly warmed to 0 ° C. and then stirred for an additional hour and then concentrated under reduced pressure. The crude product was taken up in Et ₂ O (530 mL), 1N aq. HCl solution (106 mL, 106 mmol) was added and the resulting biphasic mixture was stirred at room temperature for 3.5 hours. The layers were separated and the aqueous phase was washed with Et ₂ O (2 ×) and then basified with saturated aq. NaHCO ₃ solution. The desired amine was extracted with Et ₂ O (3 ×), then the combined organic extracts were washed with brine, dried (MgSO ₄ ) and then concentrated under reduced pressure to give the free amine. This material was treated with 4N HCl solution in dioxane (100 mL, 400 mmol) and the same material obtained from Method A (1R, 2S) / () except that a small amount of unidentified aromatic impurity (8%) was present. 1S, 2R) -1-amino-2-vinylcyclopropanecarboxylic acid ethyl ester / hydrochloric acid was obtained as a brown semi-solid (5.3 g, 34% (yield)).
Method C
C.1 Resolution of N-Boc- (1R, 2S) / (1S, 2R) -1-amino-2-vinylcyclopropanecarboxylic acid ethyl ester

分割A
39℃に維持し300rpmで撹拌している12Lジャケット(jacked)リアクター中に入れたリン酸ナトリウム緩衝剤の水性溶液(0.1M、4.25リットル(「L」)、pH8)に511グラムのAlcalase 2.4L (約425mL) (Novozymes North America Inc.)を加えた。混合物の温度が39℃に達したとき、水中の50% NaOHを加えてpHを8.0に調整した。次いで、850mLのDMSO中のラセミックN-Boc-(1R,2S)/(1S,2R)-1-アミノ-2-ビニルシクロプロパンカルボン酸エチルエステル(85g)の溶液を40分間かけて加えた。次に、反応温度を24.5時間40℃に維持し、その時間中1.5時間および19.5時間の時点で混合物のpHを水中の50% NaOHを用いて8.0に調整した。24.5時間後、エステルのエナンチオ過剰は97.2%であると測定され、次いで反応物を室温に冷却し(26℃)、一夜撹拌し(16h)、次いでエステルのエナンチオ過剰が100%であると決定された。次に、反応混合物のpHを50% NaOHで8.5に調整し、得られた混合物をMTBE (2x2L)で抽出した。次に混合MTBE抽出物を5% NaHCO₃ (3x100mL)、水(3x100mL)で洗浄し、次いで減圧下で蒸発させてエナンチオマー的に純粋なN-Boc-(1R,2S)/-1-アミノ-2-ビニルシクロプロパンカルボン酸エチルエステルを淡黄色固体として得た(42.55g；純度：97% @ 210nm、酸不含；100% エナンチオマー過剰(「ee」))。

Split A
511 grams of Alcalase 2.4L in an aqueous solution of sodium phosphate buffer (0.1M, 4.25 liters (`` L ''), pH 8) in a 12L jacked reactor maintained at 39 ° C. and stirred at 300 rpm (About 425 mL) (Novozymes North America Inc.) was added. When the temperature of the mixture reached 39 ° C., the pH was adjusted to 8.0 by adding 50% NaOH in water. A solution of racemic N-Boc- (1R, 2S) / (1S, 2R) -1-amino-2-vinylcyclopropanecarboxylic acid ethyl ester (85 g) in 850 mL DMSO was then added over 40 minutes. The reaction temperature was then maintained at 40 ° C. for 24.5 hours, at which point the pH of the mixture was adjusted to 8.0 using 50% NaOH in water at 1.5 and 19.5 hours. After 24.5 hours, the ester enantiomeric excess was measured to be 97.2%, then the reaction was cooled to room temperature (26 ° C.) and stirred overnight (16 h), then the ester enantiomeric excess was determined to be 100%. It was. The pH of the reaction mixture was then adjusted to 8.5 with 50% NaOH and the resulting mixture was extracted with MTBE (2 × 2 L). The mixed MTBE extract was then washed with 5% NaHCO ₃ (3 × 100 mL), water (3 × 100 mL) and then evaporated under reduced pressure to give enantiomerically pure N-Boc- (1R, 2S) /-1-amino- 2-Vinylcyclopropanecarboxylic acid ethyl ester was obtained as a pale yellow solid (42.55 g; purity: 97% @ 210 nm, acid free; 100% enantiomeric excess (“ee”)).

The aqueous layer from the extracted product was then acidified with 50% H ₂ SO ₄ to pH 2 and then extracted with MTBE (2 × 2 L). The MTBE extract was washed with water (3 × 100 mL) and then evaporated to give a light yellow solid (42.74 g; purity: 99% @ 210 nm, no ester).

Split B

0.5 mL 100 mM HepsNa buffer (pH 8.5), 0.1 mL Savinase 16.0 L (Protease from Bacillus clausii) (Novozymes North America Inc.) and 0.1 mL in wells of a 24-well plate (maximum volume: 10 ml / well) A solution of racemic N-Boc- (1R, 2S) / (1S, 2R) -1-amino-2-vinylcyclopropanecarboxylic acid ethyl ester (10 mg) in DMSO was added. The plate was sealed and incubated at 40 ° C., 250 rpm. After 18 hours, the enantiomeric excess of the ester was determined to be 44.3% as follows: 0.1 mL of the reaction mixture was removed and mixed well with 1 mL ethanol; after centrifugation, 10 microliters (“μL”) supernatant was removed. Analyzed by chiral HPLC. 0.1 mL DMSO was added to the remaining reaction mixture and the plates were incubated for an additional 3 days at 40 ° C., 250 rpm, then 4 mL ethanol was added to the wells. After centrifugation, 10 μL of the supernatant was analyzed by chiral HPLC and the ester enantiomeric excess was determined to be 100%.
Split C

  0.5 mL 100 mM HepsNa buffer (pH 8.5), 0.1 mL Esperase 8.0 L (Bacillus halodurans-derived protease) (Novozymes North America Inc.) and 0.1 mL in wells of a 24-well plate (maximum volume: 10 ml / well) A solution of racemic N-Boc- (1R, 2S) / (1S, 2R) -1-amino-2-vinylcyclopropanecarboxylic acid ethyl ester (10 mg) in DMSO was added. The plate was sealed and incubated at 40 ° C., 250 rpm. After 18 hours, the enantiomeric excess of the ester was determined to be 39.6% as follows: 0.1 mL of the reaction mixture was removed and mixed well with 1 mL ethanol; after centrifugation, 10 μL of the supernatant was analyzed by chiral HPLC . 0.1 mL DMSO was added to the remaining reaction mixture and the plates were incubated for an additional 3 days at 40 ° C., 250 rpm, then 4 mL ethanol was added to the wells. After centrifugation, 10 μL of the supernatant was analyzed by chiral HPLC and the ester enantiomeric excess was determined to be 100%.

Sample analysis was performed in the following manner:
1) Sample preparation: About 0.5 ml of the reaction mixture was mixed well with 10 volumes of EtOH. After centrifugation, 10 μl of supernatant was injected onto the HPLC column.
2) Conversion measurement:
Column: YMC ODS A, 4.6x50mm, S-5μm
Solvent: A, 1 mM HCl, water; B, MeCN
Gradient: 30% B, 1 minute; 30% to 45% B, 0.5 minute; 45% B, 1.5 minute; 45% to 30% B, 0.5 minute.
Flow rate: 2ml / min
UV detection: 210nm
Retention time: acid, 1.2 min; ester, 2.8 min.
3) Enantio-excess measurement of ester:
Column: CHIRACEL OD-RH, 4.6x150mm, S-5μm
Mobile phase: MeCN / 50 mM HClO ₄ , underwater (67/33)
Flow rate: 0.75ml / min.
UV detection: 210nm.
Retention time:
(1S, 2R) -1-Amino-2-vinylcyclopropanecarboxylic acid, 5.2 min;
Racemate (1R, 2S) / (1S, 2R) -1-amino-2-vinylcyclopropanecarboxylic acid ethyl ester, 18.5 min and 20.0 min;
(1R, 2S) -1-Amino-2-vinylcyclopropanecarboxylic acid ethyl ester, 18.5 min.
Split D

5 L of 0.3 M sodium phosphate buffer (pH 8) was maintained at 38 ° C. in a 20 L jacket reactor and stirred at 130 rpm. 4L Alcalase 2.4L (Novozymes North America Inc.) and 1L DI water were added to the reactor. When the temperature of the mixture reached approximately 38 ° C., the pH was adjusted to 7.8 with 10 N NaOH. A solution of racemic N-Boc- (1R, 2S) / (1S, 2R) -1-amino-2-vinylcyclopropanecarboxylic acid ethyl ester (500 g) in 5 L of DMSO was added to the addition funnel over 1 hour. Added to the reactor. The reaction temperature was then adjusted to 48 ° C. After 21 hours, the enantiomeric excess of the ester reached 99.3%. Heating was stopped at 24 hours and the reaction was gradually cooled to room temperature (about 25 ° C.) and stirred overnight. The pH of the reaction mixture was adjusted to 8.5 with 10 N NaOH and the mixture was extracted with MTBE (2 × 4 L). The combined MTBE extract was washed with 5% NaHCO ₃ ( ₃ × 400 ml) and water ( ₃ × 400 ml) and then evaporated to give enantiomerically pure N-Boc- (1R, 2S) / 1-amino-2-vinylcyclopropane The carboxylic acid ethyl ester was obtained as pale yellow crystals (259 g; purity: 96.9% @ 210 nm, acid free; 100% ee).
Split E

10 L of 0.1 M sodium phosphate buffer (pH 8) was maintained at 40 ° C. in a 20 L jacket reactor and stirred at 360 rpm. 1.5L Alcalase 2.4L (Novozymes North America Inc.) was added to the reactor. When the temperature of the mixture reached approximately 38 ° C., the pH was adjusted to 8.0 with 10N NaOH. A solution of racemic N-Boc- (1R, 2S) / (1S, 2R) -1-amino-2-vinylcyclopropanecarboxylic acid ethyl ester (200 g) in 2 L DMSO was added to the reactor from the addition funnel over 1 hour. Added to. Next, the reaction temperature was adjusted to 40 ° C. After 3 hours, the pH was adjusted to 8.0 with 10N NaOH. After 21 hours, the reaction was cooled to 25 ° C. The pH of the reaction mixture was adjusted to 8.5 with 10N NaOH and the mixture was extracted with MTBE (2 × 5 L). The combined MTBE extract was washed with 5% NaHCO ₃ ( ₃ × 500 ml) and water ( ₃ × 200 ml) and then evaporated to give 110 g of a yellow oil. The oil was set at room temperature under house vacuum to give enantiomerically pure N-Boc- (1R, 2S) / 1-amino-2-vinylcyclopropanecarboxylic acid ethyl ester as colorless long bar crystals. 101 g; purity: 97.9% @ 210 nm, acid free; 100% ee).

The crystal structure of enantiomerically pure N-Boc- (1R, 2S) /-1-amino-2-vinylcyclopropanecarboxylic acid ethyl ester was characterized by single crystal analysis (X-ray NB #: 52795-093, ref code: 643992N1). The absolute configuration is not established because there are no known chiral centers or heavier atoms. A chain structure along the crystallographic a-axis is formed via an intermolecular hydrogen bond between the amide group and the carbonyl oxygen atom (N ... O 3.159 Å).
Structure of N-Boc- (1R, 2S) -1-amino-2-vinylcyclopropanecarboxylic acid ethyl ester:

Split F

5 L of 0.2 M sodium borate buffer (pH 9) was maintained at 45 ° C. in a 20 L jacket reactor and stirred at 400 rpm. 3 L DI water and 4 L Savinase 16 L, type EX (Novozymes North America Inc.) were added to the reactor. When the temperature of the mixture was approximately 45 ° C., the pH was adjusted to 8.5 with 10 N NaOH. A solution of racemic N-Boc- (1R, 2S) / (1S, 2R) -1-amino-2-vinylcyclopropanecarboxylic acid ethyl ester (200 g) in 2 L DMSO was added to the reactor from the addition funnel over 40 minutes. Added to. Next, the reaction temperature was adjusted to 48 ° C. After 2 hours, the pH was adjusted to pH 9.0 with 10N NaOH. After 18 hours, the enantiomeric excess of the ester reached 72% and the pH was adjusted to 9.0 with 10N NaOH. After 24 hours, the temperature was lowered to 35 ° C. After 42 hours, the temperature was raised to 48 ° C. and the pH was adjusted to 9.0 with 10N NaOH. Heating was stopped at 48 hours and the reaction was gradually cooled to room temperature (about 25 ° C.) and stirred overnight. After 66 hours, the pH of the reaction mixture was 8.6. The mixture was extracted with MTBE (2x4L). The combined MTBE extract was washed with 5% NaHCO ₃ (6 × 300 ml) and water ( ₃ × 300 ml) and then evaporated to give enantiomerically pure N-Boc- (1R, 2S) / 1-amino-2-vinylcyclopropane Carboxylic acid ethyl ester was obtained as pale yellow crystals (101 Ag; purity: 95.9% @ 210 nm, acid free; 98.6% ee).
C.2 Preparation of chiral (1R, 2S) -1-amino-2-vinylcyclopropanecarboxylic acid ethyl ester / hydrochloric acid

N-BOC- (1R, 2S) -1-amino-2-vinylcyclopropanecarboxylic acid ethyl ester (8.5 g, 33.3 mmol) under N ₂ atmosphere with 200 mL 4N HCl / dioxane (Aldrich) for 3 hours at room temperature Stir. The solvent was removed under reduced pressure keeping the temperature below 40 ° C. This gave 6.57 g (˜100%) of (1R, 2S) -1-amino-2-vinylcyclopropanecarboxylic acid ethyl ester / hydrochloric acid as a pale tan solid. ¹ H NMR (300 MHz, CD ₃ OD) δ 1.31 (t, J = 7.0 Hz, 3H), 1.69-1.82 (m, 2H), 2.38 (q, J = 8.8 Hz, 1H), 4.29 (q, J = 7.0Hz, 2H), 5.22 (d, J = 10.3Hz, 1H), 5.40 (d, J = 17.2Hz, 1H), 5.69-5.81 (m, 1H). LC-MS (Method A, retention time: 0.58) min), MS m / z 156 (M ⁺ +1).
Example 2 Compound 1, 14-tert-butoxycarbonylamino-18- (isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-ene -4-carboxylic acid production

工程2A、2-(1-エトキシカルボニル-2-ビニル-シクロプロピルカルバモイル)-4-(イソキノリン-1-イルオキシ)-ピロリジン-1-カルボン酸tert-ブチルエステル

Step 2A , 2- (1-Ethoxycarbonyl-2-vinyl-cyclopropylcarbamoyl) -4- (isoquinolin-1-yloxy) -pyrrolidine-1-carboxylic acid tert-butyl ester

A stirred slurry of 4- (isoquinolin-1-yloxy) -pyrrolidine-1,2-dicarboxylic acid 1-tert-butyl ester (5.69 g, 15.9 mmol) in 200 mL methylene chloride was added to diisopropylethylamine (13.8 mL, 79.0 mmol). ), HBTU (7.10 g, 18.7 mmol), HOBTH ₂ O (2.86 g, 18.7 mmol), and 1R, 2S-1-amino-2-vinylcyclopropanecarboxylic acid ethyl ester / hydrochloric acid (3.19 g, 16.7 mmol) Processed sequentially. The golden homogeneous solution was stirred at room temperature under N ₂ for 18 hours and then concentrated under reduced pressure to give 10 g of a brown oil. This was partitioned between ethyl acetate and sat. Aq. NaHCO ₃ . The organic layer was washed with brine, dried (MgSO ₄ ) and then concentrated under reduced pressure. 5.5 g (70%) 2- (1-ethoxycarbonyl-2-vinyl-cyclopropylcarbamoyl) -4- (isoquinolin-1-yloxy) -pyrrolidine- by flash chromatography (2-5% MeOH in methylene chloride) 1-carboxylic acid tert-butyl ester was obtained: LC-MS (Method A, retention time: 3.42 min), MS m / z 496 (M ⁺ +1).
Step 2B , Preparation of 1-{[4- (Isoquinolin-1-yloxy) -pyrrolidine-2-carbonyl] -amino} -2-vinyl-cyclopropanecarboxylic acid ethyl ester, bishydrochloric acid

A stirred slurry of 2- (1-ethoxycarbonyl-2-vinyl-cyclopropylcarbamoyl) -4- (isoquinolin-1-yloxy) -pyrrolidine-1-carboxylic acid tert-butyl ester (5.40 g, 10.9 mmol) was added to 2N Treated with HCl / ether (Aldrich) (250 mL) for 24 h. The reaction mixture was concentrated under reduced pressure and 5.3 g (˜100%) of ethyl 1-{[4- (isoquinolin-1-yloxy) -pyrrolidine-2-carbonyl] -amino} -2-vinyl-cyclopropanecarboxylate The ester, bis-hydrochloric acid was obtained as a white solid: LC-MS (Method A, retention time: 2.40 min), MS m / z 396 (M ⁺ +1).
Step 2C , 1-{[1- (2-tert-Butoxycarbonylamino-nona-8-enoyl) -4- (isoquinolin-1-yloxy) -pyrrolidin-2-carbonyl] -amino} -2-vinyl-cyclo Propanecarboxylic acid ethyl ester production

2 (S) -tert-butoxycarbonylamino-8-nonenoic acid (purchased from RSP Amino Acids) (1.0 g, 3.68 mmol) dissolved in 100 mL of DMF was sequentially added to 1-{[4- (isoquinolin-1-yloxy ) -Pyrrolidine-2-carbonyl] -amino} -2-vinyl-cyclopropanecarboxylic acid ethyl ester, bishydrochloric acid (1.46 g, 3.68 mmol), N-methylmorpholine (1.4 mL, 12.9 mmol), and HATU (PE biosystems ) (1.68 g, 4.42 mmol). The reaction mixture was stirred at room temperature for 24 h under N ₂ and then concentrated under reduced pressure. The residue was partitioned between ethyl acetate and pH 4 buffer (biphthalate). The organic layer was washed with sat. Aq. NaHCO ₃ , dried (MgSO ₄ ) and then concentrated under reduced pressure to give 2.2 g of crude product. 1.9 g (79%) of 1-{[1- (2-tert-butoxycarbonylamino-nona-8-enoyl) -4- (isoquinolin-1-yloxy) by flash chromatography (20-50% ethyl acetate / hexane) ) -Pyrrolidine-2-carbonyl] -amino} -2-vinyl-cyclopropanecarboxylic acid ethyl ester was obtained as a glassy colorless solid: LC-MS (Method A, retention time: 3.72 min), MS m / z 649 (M ⁺ +1).
Step 2D , 14-tert-butoxycarbonylamino-18- (isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-en-4- Production of carboxylic acid ethyl ester

1-{[1- (2-tert-Butoxycarbonylamino-nona-8-enoyl) -4- (isoquinolin-1-yloxy) -pyrrolidine-2-carbonyl] -amino} -2 in 650 mL of methylene chloride -Vinyl-cyclopropanecarboxylic acid ethyl ester (1.72 g, 2.65 mmol) and tricyclohexylphosphine [1,3-bis (2,4,6-trimethylphenyl) -4,5-dihydroimidazol-2-ylidene] [benzylidene ] ruthenium (IV) dichloride (Strem) (225mg, 0.265mmol) was refluxed under N _2. The pale orange homogeneous solution was refluxed for 24 hours to give a dark orange solution. The reaction mixture was cooled to room temperature, then cooled to room temperature and then concentrated under reduced pressure to give 1.7 g of an orange oil. By flash chromatography (20-40% ethyl acetate / hexane) 1.4 g (85%) 14-tert-butoxycarbonylamino-18- (isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza- Tricyclo [14.3.0.0 ^4,6 ] -nonadeca-7-ene-4-carboxylic acid ethyl ester was obtained as a white solid: ¹ H NMR (300 MHz, CD ₃ Cl ₃ ) δ 1.29 (t, J = 7.3 Hz, 3H), 1.34 (s, 9H), 1.35-1.71 (m, 9H), 1.86-1.97 (m, 2H), 2.08-2.27 (m, 2H), 2.38-2.47 (m, 1H), 2.96-3.04 ( m, 1H), 4.02-4.23 (m, 4H), 4.55 (m, 1H), 4.91 (dd, J = 8.4Hz, 4.0Hz, 1H), 5.25 (t, J = 9.5Hz, 1H), 5.32 ( d, J = 8.8Hz, 1H), 5.50 (m, 1H), 5.81 (m, 1H), 6.96 (s, 1H), 7.23 (m, 1H), 7.51 (t, J = 7.3Hz, 1H), 7.65 (t, J = 8.4Hz, 1H), 7.73 (d, J = 8.4Hz, 1H), 7.93 (d, J = 5.9Hz, 1H), 8.17 (d, J = 8.4Hz, 1H). LC- MS (Method A, retention time: 3.52 min), MS m / z 621 (M ⁺ +1).
Step 2E , Compound 1, 14-tert-butoxycarbonylamino-18- (isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-ene -4-carboxylic acid production

14-tert-Butoxycarbonylamino-18- (isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] -nonadeca-7-ene-4-carboxylic acid The ethyl ester (1.10 g, 1.77 mmol) was dissolved in a mixed solvent system: THF (18 mL), methanol (8 mL), and water (2 mL). Powdered lithium hydroxide hydrate (744 mg, 17.7 mmol) was added. The pale yellow slurry was stirred at room temperature for 24 hours under N ₂ and then concentrated under reduced pressure. The residue was partitioned between ether and water. The ether phase was discarded and the aqueous phase was treated with 4N HCl until the pH was 4. The acidic solution was extracted with ether for 4 hours. The mixed ether extract was dried (MgSO ₄ ) and then concentrated under reduced pressure to give 0.95 g (90%) of 14-tert-butoxycarbonylamino-18- (isoquinolin-1-yloxy) -2,15-dioxo- 3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] -nonadeca-7-ene-4-carboxylic acid was obtained as a white solid. ¹ H NMR (300 MHz, CD ₃ Cl ₃ ) δ 1.30 (s, 9H), 1.34-1.46 (m, 4H), 1.52-1.68 (m, 2H), 1.84 (m, 2H), 2.08-2.58 (m, 6H), 2.78-2.87 (m, 1H), 4.07 (m, 1H), 4.32 (d, J = 11.3Hz, 1H), 4.41 (m, 1H), 4.79 (t, J = 7.3Hz, 1H), 5.14 (t, J = 9.5Hz, 1H), 5.23 (d, J = 7.3Hz, 1H), 5.60 (q, J = 8.8Hz, 1H), 5.86 (s, 1H), 7.08 (s, 1H), 7.22 (d, J = 6.2Hz, 1H), 7.48 (t, J = 7.7Hz, 1H), 7.64 (t, J = 8.0Hz, 1H), 7.72 (d, J = 8.1Hz, 1H), 7.94 ( d, J = 5.9 Hz, 1H), 8.17 (s, J = 8.4 Hz, 1H). LC-MS (Method A, retention time: 3.32 min), MS m / z 593 (M ⁺ +1).
Example 3a , Preparation of cyclopropylsulfonamide
Method A :

Gaseous ammonia was bubbled through a solution of 100 mL of THF cooled to 0 ° C. until saturation was reached. To this solution was added a solution of 5 g (28.45 mmol) cyclopropylsulfonyl chloride (purchased from Array Biopharma) in 50 mL THF and the solution was allowed to warm to room temperature overnight and stirred for an additional day. The mixture is concentrated until _1-2 mL of solvent remains, then applied to a plug of 30 g SiO2 (eluting with 30% -60% EtOAc / hexanes) 3.45 g (100%) of cyclopropylsulfonamide as a white solid Got as. ¹ H NMR (methanol-d ₄ ) δ 0.94-1.07 (m, 4H), 2.52-2.60 (m, 1H); ¹³ C NMR (methanol-d4) δ 5.92, 33.01.
Method B :
Step 1 : Production of N-tert-butyl- (3-chloro) propylsulfonamide

tert-Butylamine (3.0 mol, 315.3 mL) was dissolved in THF (2.5 L). The solution was cooled to -20 ° C. 3-Chloropropanesulfonyl chloride (1.5 mol, 182.4 mL) was added slowly. The reaction mixture was warmed to room temperature and stirred for 24 hours. The mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was dissolved in CH ₂ Cl ₂ (2.0 L). The resulting solvent was washed with 1N HCl (1.0 L), water (1.0 L), brine (1.0 L) and then dried over Na ₂ SO ₄ . It was filtered and then concentrated under reduced pressure to give a pale yellow solid, which was crystallized from hexane to give a white solid product (316.0 g, 99%).
¹ H NMR (CDCl ₃ ) δ 1.38 (s, 9H), 2.30-2.27 (m, 2H), 3.22 (t, J = 7.35Hz, 2H), 3.68 (t, J = 6.2Hz, 2H), 4.35 ( b, 1H).
Step 2 : Production of cyclopropanesulfonic acid tert-butylamide

To a solution of N-tert-butyl- (3-chloro) propylsulfonamide (2.14 g, 10.0 mmol) in THF (100 mL) at −78 ° C. n-BuLi (2.5 M in hexane, 8.0 mL, 20.0 mmol) Was added. The reaction mixture was allowed to warm to room temperature over 1 hour. Volatiles were removed under reduced pressure. The residue was partitioned between EtOAC and water (200 mL, 200 mL). The separated organic phase was washed with brine, dried (Na ₂ SO ₄ ), filtered and then concentrated under reduced pressure. The residue was recrystallized from hexanes to give the desired product as a white solid (1.0 g, 56%).
¹ H NMR (CDCl ₃ ) δ 0.98-1.00 (m, 2H), 1.18-1.19 (m, 2H), 1.39 (s, 9H), 2.48-2.51 (m, 1H), 4.19 (b, 1H).
Process 3 : Production of cyclopropylsulfonamide

A solution of cyclopropanesulfonic acid tert-butylamide (110.0 g, 0.62 mol) in TFA (500 mL) was stirred at room temperature for 16 hours. Volatiles were removed under reduced pressure. The residue was recrystallized from EtOAC / hexane (60 mL / 240 mL) to give the desired product as a white solid (68.5 g, 91%).
¹ H NMR (DMSO-d ₆ ) δ 0.84-0.88 (m, 2H), 0.95-0.98 (m, 2H), 2.41-2.58 (m, 1H), 6.56 (b, 2H).
Example 3b Preparation of C1-substituted cyclopropylsulfonamide
Preparation of N-tert-butyl- (1-methyl) cyclopropyl-sulfonamide

工程1a：N-tert-ブチル-(3-クロロ)プロピルスルホンアミドの製造

Step 1a : Production of N-tert-butyl- (3-chloro) propylsulfonamide

As above.
Step 1b : Preparation of N-tert-butyl- (1-methyl) cyclopropyl-sulfonamide

A solution of N-tert-butyl- (3-chloro) propylsulfonamide (4.3 g, 20 mmol) was dissolved in dry THF (100 mL) and cooled to -78 ° C. To this solution was added n-BuLi (17.6 mL, 44 mmol, 2.5 M in hexane) gradually. The dry ice bath was removed and the reaction mixture was warmed to room temperature for 1.5 hours. The mixture was then cooled to −78 ° C. and then n-BuLi solution (20 mmol, 8 mL, 2.5 M in hexane) was added. The reaction mixture was allowed to warm to room temperature and then re-cooled to −78 ° C. over 2 hours and a raw solution of methyl iodide (5.68 g, 40 mmol) was added. The reaction mixture was warmed to room temperature overnight and then quenched with saturated NH ₄ Cl (100 mL) at room temperature. It was extracted with EtOAc (100 mL). The organic layer was washed with brine (100 mL), dried (MgSO ₄ ) and then concentrated under reduced pressure to give a yellow oil which crystallized from hexane to give the product as a pale yellow solid (3.1 g , 81%): ¹ H NMR (CDCl ₃ ) δ 0.79 (m, 2H), 1.36 (s, 9H), 1.52 (m, 2H), 1.62 (s, 3H), 4.10 (bs, 1H).
Step 1c : Production of 1-methylcyclopropylsulfonamide

A solution of N-tert-butyl- (1-methyl) cyclopropylsulfonamide (1.91 g, 10 mmol) was dissolved in TFA (30 mL) and the reaction mixture was stirred at room temperature for 16 hours. The solvent was removed under reduced pressure to give a yellow oil that was crystallized from EtOAc / hexane (1: 4, 40 mL) to give Example 3, 1-methylcyclopropylsulfonamide as a white solid (1.25 g). 96%): ¹ H NMR (CDCl ₃ ) δ 0.84 (m, 2H), 1.41 (m, 2H), 1.58 (s, 3H), 4.65 (bs, 2H). Theoretical value of _{C 4 H 9 NO 2 S:} C, 35.54; H, 6.71; N, 10.36. Found: C, 35.67; H, 6.80; N, 10.40.
Production of 1-benzylcyclopropylsulfonamide

工程1b：N-tert-ブチル-(1-ベンジル)シクロプロピル-スルホンアミドの製造

Step 1b : Preparation of N-tert-butyl- (1-benzyl) cyclopropyl-sulfonamide

This compound was collected using the method described for the synthesis of N-tert-butyl- (1-methyl) cyclopropylsulfonamide except that 1.05 equivalents of benzyl bromide was then triturated with 10% EtOAc in hexane. Obtained at a rate of 60%: ¹ H NMR (CDCl ₃ ) δ 0.92 (m, 2H), 1.36 (m, 2H), 1.43 (s, 9H), 3.25 (s, 2H), 4.62 (bs, 1H), 7.29-7.36 (m, 5H).
Step 1c : Preparation of 1-benzylcyclo-propylsulfonamide

This compound, 1-benzylcyclopropylsulfonamide, was recrystallized using the method described for the synthesis of 1-methylcyclopropylsulfonamide and then from a minimum amount of 10% EtOAc in hexanes to give N-tert-butyl (1- Obtained from benzyl) cyclopropylsulfonamide in 66% yield: ¹ H NMR (CDCl ₃ ) δ 0.90 (m, 2H), 1.42 (m, 2H), 3.25 (s, 2H), 4.05 (s, 2H) , 7.29 (m, 3H), 7.34 (m, 2 H); 13 C NMR (CDCl 3) δ 11.1,36.8,41.9,127.4,128.8,129.9,136.5.
Production of 1-propylcyclopropylsulfonamide

工程1b：N-tert-ブチル-(1-ベンジル)シクロプロピル-スルホンアミドの製造

Step 1b : Preparation of N-tert-butyl- (1-benzyl) cyclopropyl-sulfonamide

This compound was prepared using the method described for the preparation of 1-methylcyclopropylsulfonamide except that propyl halide was used in place of methyl iodide in the second step of the method.
Production of N-tert-butyl- (1-allyl) cyclopropylsulfonamide

This compound, N-tert-butyl- (1-allyl) cyclopropylsulfonamide, was used except that 1.25 equivalents of allyl bromide was used as the electrophile. The yield was 97% according to the method described in the synthesis. The compound was used directly in the next reaction without further purification: ¹ H NMR (CDCl ₃ ) δ 0.83 (m, 2H), 1.34 (s, 9H), 1.37 (m, 2H), 2.64 (d, J = 7.3 Hz, 2H), 4.25 (bs, 1H), 5.07-5.10 (m, 2H), 6.70-6.85 (m, 1H).
Production of 1-allylcyclopropylsulfonamide

This compound, 1-allylcyclopropylsulfonamide, was obtained in 40% yield from N-tert-butyl- (1-allyl) cyclopropylsulfonamide according to the method described in the synthesis of 1-methylcyclopropylsulfonamide. The compound was purified by SiO ₂ column chromatography using 2% MeOH in CH ₂ Cl ₂ as eluent: ¹ H NMR (CDCl ₃ ) δ 0.88 (m, 2H), 1.37 (m, 2H), 2.66 (d, J = 7.0 Hz, 2H), 4.80 (s, 2H), 5.16 (m, 2H), 5.82 (m, 1H); ¹³ C NMR (CDCl ₃ ) δ 11.2, 35.6, 40.7, 119.0, 133.6.
Preparation of N-tert-butyl- [1- (1-hydroxy) cyclohexyl] -cyclopropylsulfonamide

The compound is recrystallized using the method described in the synthesis of N-tert-butyl- (1-methyl) cyclopropylsulfonamide except using 1.30 equivalents of cyclohexanone and then from a minimum amount of 20% EtOAc in hexane. 84% yield: ¹ H NMR (CDCl ₃ ) δ 1.05 (m, 4H), 1.26 (m, 2H), 1.37 (s, 9H), 1.57-1.59 (m, 6H), 1.97 (m , 2H), 2.87 (bs, 1H), 4.55 (bs, 1H).
Preparation of 1- (1-cyclohexenyl) cyclopropyl-sulfonamide

This compound, 1- (1-cyclohexenyl) -cyclopropylsulfonamide, was recrystallized from the minimum amount of EtOAc and hexanes using the method described for the synthesis of 1-methylcyclopropylsulfonamide and then N-tert- Obtained from butyl- [1- (1-hydroxy) cyclohexyl] -cyclopropylsulfonamide in 85% yield: ¹ H NMR (DMSO-d6) δ 0.82 (m, 2H), 1.28 (m, 2H), 1.51 (m, 2H), 1.55 (m, 2H), 2.01 (s, 2H), 2.16 (s, 2H), 5.89 (s, 1H), 6.46 (s, 2H); ¹³ C NMR (DMSO-d ₆ ) 11.6, 21.5, 22.3, 25.0, 27.2, 46.9, 131.6, 132.2; LR-MS (ESI): 200 (M ⁺ -1).
Preparation of N-tert-butyl- (1-benzoyl) cyclopropyl-sulfonamide

This compound was obtained in 66% yield using the method described for the synthesis of N-tert-butyl- (1-methyl) cyclopropylsulfonamide except that 1.2 equivalents of methyl benzoate was used as electrophile. The compound was purified by SiO ₂ column chromatography using 30% to 100% CH ₂ Cl ₂ in hexane: ¹ H NMR (CDCl ₃ ) δ 1.31 (s, 9H), 1.52 (m, 2H), 1.81 (m, 2H), 4.16 (bs, 1H), 7.46 (m, 2H), 7.57 (m, 1H), 8.05 (d, J = 8.5Hz, 2H).
Production of 1-benzoylcyclo-propylsulfonamide

This compound, 1-benzoylcyclopropyl-sulfonamide, was recrystallized from the minimum amount of EtOAc in hexane using the method described for the synthesis of 1-methylcyclopropylsulfonamide and then N-tert-butyl (1-benzoyl ) Obtained from cyclopropylsulfonamide in 87% yield: ¹ H NMR (DMSO-d ₆ ) δ 1.39 (m, 2H), 1.61 (m, 2H), 7.22 (s, 2H), 7.53 (t, J = 7.6 Hz, 2H), 7.65 (t, J = 7.6 Hz, 1H), 8.06 (d, J = 8.2 Hz, 2 H); ¹³ C NMR (DMSO-d ₆ ) δ 12.3, 48.4, 128.1, 130.0, 133.4, 135.3, 192.0.
Preparation of N-tert-butyl- (1-phenylaminocarboxy) -cyclopropylsulfonamide

This compound is recrystallized from 1 min of phenylisocyanate using the method described for the synthesis of N-tert-butyl- (1-methyl) cyclopropylsulfonamide and then recrystallized from a minimum amount of EtOAc in hexane to yield. Obtained at 42%. ¹ H NMR (CDCl ₃ ) δ 1.38 (s, 9H), 1.67-1.71 (m, 4H), 4.30 (bs, 1H), 7.10 (t, J = 7.5Hz, 1H), 7.34 (t, J = 7.5 Hz, 2H), 7.53 (t, J = 7.5Hz, 2H).
Example 4 Preparation of cycloalkylsulfonamide from cycloalkylbromide
Production of cyclobutylsulfonamide from cyclobutyl bromide

To a solution of 5.0 g (37.0 mmol) cyclobutyl bromide in 30 mL anhydrous diethyl ether (Et ₂ O) cooled to −78 ° C. was added 44 mL (74.8 mmol) 1.7 M tert-butyllithium in pentane, then The solution was gradually warmed to -35 ° C over 1.5 hours. This mixture was slowly cannulated into a solution of 5.0 g (37.0 mmol) of freshly distilled sulfuryl chloride in 100 mL hexane cooled to -40 ° C., warmed to 0 ° C. over 1 hour and then carefully concentrated under reduced pressure. . This mixture was redissolved in Et ₂ O, washed once with some ice-cold water, dried (MgSO ₄ ) and then carefully concentrated. This mixture was redissolved in 20 mL of THF and added dropwise to 500 mL of saturated NH _{3 in} THF and then stirred overnight. The mixture was concentrated under reduced pressure to give a crude yellow solid that was recrystallized from CH ₂ Cl _{2 in} hexane containing a minimum amount of 1-2 drops of MeOH to give 1.90 g (38%) of cyclobutylsulfonamide. Obtained as a white solid. ¹ H NMR (CDCl ₃ ) δ 1.95-2.06 (m, 2H), 2.30-2.54 (m, 4H), 3.86 (p, J = 8 Hz, 1H), 4.75 (brs, 2H); ¹³ C NMR (CDCl ₃ ) δ 16.43, 23.93, 56.29. HRMS m / z (MH) ⁻ , Theoretical value for C ₄ H ₈ NSO ₂ : 134.0276, Found: 134.0282.
Production of cyclopentylsulfonamide

A solution of 18.5 mL (37.0 mmol) of 2M cyclopentyl-magnesium chloride in ether was added dropwise to a solution of 3.0 mL (37.0 mmol) freshly distilled sulfuryl chloride (obtained from Aldrich) in 100 mL hexane cooled to -78 ° C. did. The mixture was warmed to 0 ° C. over 1 h and then carefully concentrated under reduced pressure. The mixture was redissolved in Et ₂ O (200 mL), washed once with some ice-cold water (200 mL), dried (MgSO ₄ ) and then carefully concentrated. This mixture was redissolved in 35 mL of THF, added dropwise to 500 mL of saturated NH _{3 in} THF and then stirred overnight. The mixture was concentrated under reduced pressure to give a crude yellow solid, the residue was filtered through 50 g of silica gel using 70% EtOAc-hexane as eluent, then the solvent was concentrated. The residue was recrystallized from CH ₂ Cl _{2 in} hexane containing a minimum amount of 1-2 drops of MeOH to give 2.49 g (41%) of cyclopentylsulfonamide as a white solid. ¹ H NMR (CDCl ₃ ) δ 1.58-1.72 (m, 2H), 1.74-1.88 (m, 2H), 1.94-2.14 (m, 4H), 3.48-3.59 (m, 1H), 4.80 (bs, 2H) ¹³ C NMR (CDCl ₃ ) δ 25.90, 28.33, 63.54; MS m / e 148 (MH) ⁻ .
Production of cyclohexylsulfonamide

A solution of 2M cyclohexyl magnesium chloride in ether (TCI Americas) in 18.5 mL (37.0 mmol) was added dropwise to a solution of 3.0 mL (37.0 mmol) freshly distilled sulfuryl chloride in 100 mL hexane cooled to -78 ° C. The mixture was warmed to 0 ° C. over 1 h and then carefully concentrated under reduced pressure. This mixture was redissolved in Et ₂ O (200 mL), washed once with some ice-cold water (200 mL), dried (MgSO ₄ ) and then carefully concentrated. This mixture was redissolved in 35 mL of THF and added dropwise to 500 mL of saturated NH ₃ in THF and then stirred overnight. The mixture was concentrated under reduced pressure to a crude yellow solid and the residue was filtered through 50 g of silica gel using 70% EtOAc-hexane as eluent and then concentrated. The residue was recrystallized from CH ₂ Cl _{2 in} hexane containing a minimum amount of 1-2 drops of MeOH to give 1.66 g (30%) of cyclohexyl-sulfonamide as a white solid: ¹ H NMR (CDCl ₃ ) δ 1.11-1.37 (m, 3H), 1.43-1.56 (m, 2H), 1.67-1.76 (m, 1H), 1.86-1.96 (m, 2H), 2.18-2.28 (m, 2H), 2.91 (tt, J = 12, 3.5 Hz, 1H), 4.70 (bs, 2H); ¹³ CH NMR (CDCl ₃ ) δ 25.04, 25.04, 26.56, 62.74; MS m / e 162 (M-1) ⁻ .
Example 5 Compound 2, [18- (isoquinolin-1-yloxy) -4-methanesulfonylaminocarbonyl-2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-ene Of -14-yl] -carbamic acid tert-butyl ester

14-tert-butoxycarbonylamino-18- (isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] -nonadeca as described in the general method above 7-ene-4-carboxylic acid (50 mg, 0.075 mmol) and methanesulfonamide (9.3 mg, 0.098 mmol) to [18- (isoquinolin-1-yloxy) -4-methanesulfonylaminocarbonyl-2,15-dioxo -3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-en-14-yl] -carbamic acid tert-butyl ester was obtained as a white powder: ¹ H NMR (300 MHz, CD ₃ Cl ₃ ) δ 1.18 (s, 9H), 1.25-1.96 (m, 11H), 2.26 (m, 1H), 2.54 (m, 1H), 2.71 (m, 2H), 3.18 (s, 3H), 4.08 (m, 1H), 4.26 (m, 1H), 4.64 (m, 2H), 5.00 (m, 2H), 5.73 (m, 1H), 6.01 (s, 1H), 6.73 (s, 1H), 7.28 (d, J = 5.5Hz, 1H), 7.50 (t, J = 7.3Hz, 1H), 7.65-7.76 (m, 2H), 7.98 (d, J = 5.9Hz, 1H), 8.19 (d, J = 7.7Hz, 1H) ), 10.28 (s, 1H). LC-MS (Method A, holding) Between: 3.57 min), MS m / z 670 (M ⁺ +1).
Example 6 Compound 3, [18- (isoquinolin-1-yloxy) -2,15-dioxo-4- (propane-2-sulfonylaminocarbonyl) -3,16-diaza-tricyclo- [14.3.0.0 ^4,6 ] Preparation of nonadeca-7-en-14-yl] -carbamic acid tert-butyl ester

14-tert-Butoxycarbonylamino-18- (isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] -nonadeca-7-ene-4-carboxylic acid (67 mg, 0.10 mmol) was dissolved in 5 mL of THF and then treated with CDI (21 mg, 0.13 mmol). (An oven-dried glass container was used, taking care to maintain a dry N ₂ atmosphere and avoid moisture.) The reaction mixture was refluxed for 1 hour, then cooled to room temperature, then isopropylsulfonamide (16 mg, 0.13 mmol) and DBU. (20 mg, 0.13 mmol). After stirring at room temperature for 24 hours, THF was removed by rotary evaporation. The residue was partitioned between ethyl acetate and pH 4 buffer. The organic layer was dried (MgSO ₄ ) and then concentrated under reduced pressure to give the crude product. Flash chromatography (1-5% MeOH / methylene chloride) gave 50 mg (71%) of the desired product. Further purification by preparative HPLC (YMC ODS-A, S5, 20 × 100 mm, gradient: 60% to 100% B) followed by 30 mg (43%) of [18- (isoquinolin-1-yloxy) -2,15-dioxo- 4- (Propane-2-sulfonylaminocarbonyl) -3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-en-14-yl] -carbamic acid tert-butyl ester was obtained as a white powder : ¹ H NMR (300 MHz, CD ₃ Cl ₃ ) δ 1.22 (S, 9H), 1.31 (d. J = 6.6 Hz, 3H), 1.41 (d, J = 6.9 Hz, 3H), 1.20-1.96 (m, 11H), 2.30 (q, J = 8.4Hz, 1H), 2.56 (m, 1H), 2.69 (m, 2H), 3.69 (m, 1H), 4.03 (m, 1H), 4.29 (m, 1H), 4.62 (m, 2H), 4.97-5.06 (m, 2H), 5.68 (q, J = 9.5Hz, 1H), 5.91 (s, 1H), 6.76 (s, 1H), 7.23 (m, 1H), 7.45 (t, J = 7.7Hz, 1H), 7.63 (t, J = 8.1Hz, 1H), 7.71 (d, J = 8.1Hz, 1H), 7.97 (d, J = 5.9Hz, 1H), 8.17 (d , J = 8.1 Hz, 1H), 9.89 (s, 1H). LC-MS (Method A, retention time: 3.79 min), MS m / z 670 (M ⁺ +1).
Example 7 Compound 4, [4-cyclopropanesulfonylaminocarbonyl-18- (isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7- En-14-yl] -carbamic acid tert-butyl ester

14-tert-butoxycarbonylamino-18- (isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] -nonadeca as described in the general method above Prepared from 7-ene-4-carboxylic acid etherate (100 mg, 0.15 mmol) and cyclopropylsulfonamide (24 mg, 0.20 mmol). Flash chromatography (2% methanol / methylene chloride) gave 55 mg (53%) of [4-cyclopropanesulfonylaminocarbonyl-18- (isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo- [14.3.0.0 ^4,6 ] -Nonadeca-7-en-14-yl] -carbamic acid tert-butyl ester was obtained as a white powder: ¹ H NMR (500 MHz, CD ₃ Cl ₃ ) δ 0.89-0.95 (m , 1H), 1.03-1.16 (m, 3H), 1.22 (s, 9H), 1.18-1.48 (m, 8H), 1.78 (m, 1H), 1.87 (m, 1H), 1.94 (m, 1H), 2.29 (q, J = 9.0Hz, 1H), 2.56 (m, 1H), 2.67 (dd, J = 7.9Hz, 2.7Hz, 2H), 2.90 (m, 1H), 4.04 (dd, J = 11.3Hz, 4.0Hz, 1H), 4.29 (m, 1H), 4.61 (m, 2H), 4.97-5.03 (m, 2H), 5.70 (m, 1H), 5.94 (s, 1H), 6.67 (s, 1H), 7.25 (m, 1H), 7.47 (t, J = 7.3Hz, 1H), 7.64 (t, J = 7.3Hz, 1H), 7.72 (d, J = 7.9Hz, 1H), 7.97 (d, J = 5.8 Hz, 1H), 8.17 (d, J = 8.5 Hz, 1H), 10.20 (s, 1H). LC-MS (Method A, retention time: 3.67 min), MS m / z 696 (M ⁺ +1).
Example 8 [4- (1-Benzyl-cyclopropanesulfonylaminocarbonyl) -18- (isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca -7-en-14-yl] -carbamic acid tert-butyl ester

14-tert-Butoxycarbonylamino-18- (isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] -nonadeca-7-ene-4-carboxylic acid (50 mg, 0.075 mmol) and 1-benzyl-cyclopropanesulfonic acid amide (21 mg, 0.098 mmol, prepared as above) and [4- (1-benzyl-cyclopropanesulfonylaminocarbonyl) -18- (isoquinoline) -1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-en-14-yl] -carbamic acid tert-butyl ester was obtained as a white powder. : ¹ H NMR (500 MHz, CD ₃ Cl ₃ ) δ 1.23 (s, 9H), 1.04-1.92 (m, 14H), 1.96 (m, 1H), 2.32 (dd, J = 17.7 Hz, 9.5 Hz, 1H) , 2.59 (m, 1H), 2.70 (m, 2H), 3.20 (d, J = 13.4Hz, 1H), 3.41 (d, J = 13.7Hz, 1H), 4.02 (m, 1H), 4.29 (m, 1H), 4.61 (m, 2H), 5.01 (d, J = 7.6Hz, 1H), 5.15 (m, 1H), 5.78 (m, 1H), 5.92 (s, 1H), 6.65 (s, 1H), 7.08 (d, J = 8.2H z, 2H), 7.26 (m, 4H), 7.46 (t, J = 7.3Hz, 1H), 7.64 (t, J = 7.3Hz, 1H), 7.72 (d, J = 8.2Hz, 1H), 7.97 ( d, J = 5.8Hz, 1H), 8.17 (d, J = 8.9Hz, 1H), 10.05 (s, 1H). LC-MS (Method A, retention time: 3.87min), MS m / z 786 (M ⁺ +1).
Example 9 Compound 6, [18- (isoquinolin-1-yloxy) -2,15-dioxo-4- (1-propyl-cyclopropanesulfonylaminocarbonyl) -3,16-diaza-tricyclo [14.3.0.0 ^{4, 6} ] Production of nonadeca-7-en-14-yl] -carbamic acid tert-butyl ester

14-tert-Butoxycarbonylamino-18- (isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] -nonadeca-7-ene-4-carboxylic acid (50 mg, 0.075 mmol) and 1-propyl-cyclopropanesulfonic acid amide (16 mg, 0.098 mmol, prepared as above) and [18- (isoquinolin-1-yloxy) -2,15-dioxo-4- (1-propyl-cyclopropanesulfonylaminocarbonyl) -3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-en-14-yl] -carbamic acid tert-butyl ester was obtained as a white powder : ¹ H NMR (300 MHz, CD ₃ Cl ₃ ) δ 0.82-0.93 (m, 5H), 1.23 (s, 9H), 1.30-1.94 (m, 17H), 2.28 (m, 1 H), 2.54 (m, 1H), 2.68 (dd, J = 8.1Hz, 3.2Hz, 2H), 4.05 (m, 1H), 4.30 (m, 1H), 4.63-4.66 (m, 2H), 4.96-5.09 (m, 2H), 5.70 (m, 1H), 5.92 (s, 1H), 6.69 (s, 1H), 7.23 (m, 1H), 7.46 (t, J = 8.1Hz, 1H), 7.63 (t, J = 8.1Hz, 1H ), 7.72 (d, J = 8.1Hz, 1H), 7.97 (d, J = 5.9Hz, 1H), 8.17 (d, J = 8.4Hz, 1H), 10.12 (s, 1H). LC-MS (Method A, retention time: 3.77min) ), MS m / z 738 (M ⁺ +1).
Example 10 Compound 7, [18- (isoquinolin-1-yloxy) -2,15-dioxo-4- (1-propyl-cyclopropanesulfonylaminocarbonyl) -3,16-diaza-tricyclo [14.3.0.0 ^{4, 6} ] Production of nonadeca-14-yl] -carbamic acid tert-butyl ester

73 mg (0.1 mmol) of [18- (isoquinolin-1-yloxy) -2,15-dioxo-4- (1-propylcyclopropanesulfonylamino-carbonyl) -3,16-diaza-tricyclohexane in 3 mL of methanol To a mixture of [14.3.0.0 ^4,6 ] nonadeca-7-en-14-yl] -carbamic acid tert-butyl ester was added 332 mg (2 mmol) of dipotassium azodicarboxylate. Glacial acetic acid (240 mg, 4 mmol) in 2 mL of methanol was added slowly over 5 hours via syringe pump. The mixture was stirred at room temperature. After 5 hours another 332 mg (2 mmol) dipotassium azodicarboxylate and 240 mg (4 mmol) glacial acetic acid were added over the course of 5 hours and then stirring was continued overnight. This cycle was repeated twice. LC-MS showed about 40% starting material still remained. The solvent was then removed on a rotary evaporator. Water was added to the residue and then the mixture was extracted with ethyl acetate for 3 hours. It was then dried over magnesium sulfate, filtered and then concentrated under reduced pressure. The resulting oil was dissolved in methanol and then purified by preparative HPLC (YMC ODS-A, S5, 20x100 mm, gradient: 80% B to 85% B, 15 min, retention 2 min, flow rate 25 mL / min) A powdery product was obtained (25 mg, 34%).
¹ H NMR (500 MHz, CD ₃ Cl ₃ ) δ 0.91 (m, 5H), 1.30 (s, 9H), 1.20-1.80 (m, 21H), 1.86-1.93 (m, 2H), 2.57 (m, 1H) 2.67 (m, 1H), 4.10 (m, 1H), 4.36 (d, J = 12.2Hz, 1H), 4.44 (t, J = 8.2Hz, 1H), 4.61 (t, J = 7.9Hz, 1H) 5.16 (d, J = 8.2Hz, 1H), 5.92 (bs, 1H), 6.58 (bs, 1H), 7.25 (m, 1H), 7.49 (t, J = 7.3Hz, 1H), 7.65 (t, J = 7.3Hz, 1H), 7.73 (d, J = 8.2Hz, 1H), 7.97 (d, J = 5.8Hz, 1H), 8.15 (d, J = 8.2Hz, 1H), 10.09 (s, 1H) LC-MS (Method A, retention time: 3.83 min), MS m / z 740 (M ⁺ +1).
Example 11 Compound 8, [18- (isoquinolin-1-yloxy) -4- (1-methyl-cyclopropanesulfonylaminocarbonyl) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^{4, 6} ] Production of nonadeca-7-en-14-yl] -carbamic acid tert-butyl ester

14-tert-Butoxycarbonylamino-18- (isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] -nonadeca-7-ene-4-carboxylic acid Prepared from etherate (105 mg, 0.177 mmol) and 1-methyl-cyclopropanesulfonic acid amide (31 mg, 0.23 mmol, prepared as above). By flash chromatography (2% methanol / methylene chloride) 73 mg (58%) of [18- (isoquinolin-1-yloxy) -4- (1-methyl-cyclopropanesulfonylaminocarbonyl) -2,15-dioxo-3, 16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-en-14-yl] -carbamic acid tert-butyl ester was obtained as a white powder: LC-MS (Method A retention time: 3.50 min), MS m / z 710 (M ⁺ +1).
Example 12 Compound 9, [4-cyclobutanesulfonylaminocarbonyl-18- (isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-ene Of -14-yl] -carbamic acid tert-butyl ester

14-tert-Butoxycarbonylamino-18- (isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] -nonadeca-7-ene-4-carboxylic acid Prepared from etherate (114 mg, 0.192 mmol) and cyclobutanesulfonic acid amide (34 mg, 0.25 mmol) as described in the general method above. By flash chromatography (2% methanol / methylene chloride) 73 mg (58%) of [18- (isoquinolin-1-yloxy) -4- (1-methyl-cyclopropanesulfonylaminocarbonyl) -2,15-dioxo-3, 16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-en-14-yl] -carbamic acid tert-butyl ester was obtained as a white powder: LC-MS (Method A, retention time: 3.55 min) , MS m / z 710 (M ⁺ +1).
Example 13 Compound 10, [4-cyclopropanesulfonylaminocarbonyl-18- (isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-14- Of yl] -carbamic acid tert-butyl ester

140 mg (0.2 mmol) [4-cyclopropanesulfonylaminocarbonyl-18- (isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^{4, in} 3 mL methanol ⁶ ] To a mixture of nonadeca-7-en-14-yl] -carbamic acid tert-butyl ester was added 656 mg (4 mmol) of dipotassium azodicarboxylate. Glacial acetic acid (480 mg, 8 mmol) in 2 mL of methanol was added slowly over 5 hours with a syringe pump. The mixture was stirred at room temperature. After 5 hours, another 656 mg (4 mmol) of dipotassium azodicarboxylate and 480 mg (8 mmol) of glacial acetic acid were added over 5 hours and then stirring was continued overnight. This cycle was repeated twice. LC-MS showed about 40% starting material still remained. The solvent was then removed on a rotary evaporator. Water was added to the residue and the mixture was extracted with ethyl acetate for 3 hours. It was then dried over magnesium sulfate, filtered and then concentrated under reduced pressure. The obtained oil was dissolved in methanol and then purified by preparative HPLC (YMC XTERRA, S5, 30x50 mm, gradient: 72% B to 78% B, 15 min, retention 2 min, flow rate 40 ml / min) to give a white powder. Product was isolated (80 mg, 57%). LC-MS (Method A, retention time: 3.59 min), MS m / z 698 (M ⁺ +1).
Example 14 Compound 11, cyclopropanesulfonic acid [14-amino-18- (isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7- En-4-carbonyl] -amide bishydrochloric acid production

[4-Cyclopropane-sulfonylaminocarbonyl-18- (isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-en-14-yl A stirred slurry of] -carbamic acid tert-butyl ester (1.20 g, 1.7 mmol) was treated with 4N HCl / dioxane (Aldrich) (10 mL) for 4 h. The reaction mixture was concentrated under reduced pressure to obtain 1.10 g (92%) of cyclopropanesulfonic acid [14-amino-18- (isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3 .0.0 ^4,6 ] nonadeca-7-ene-4-carbonyl] -amide bishydrochloric acid was obtained as a white solid: LC-MS (Method B, retention time: 1.39 min), MS m / z 596 (M ⁺ + 1-2 HCl).
Example 15 Compound 12 [4-Cyclopropanesulfonylaminocarbonyl-18- (isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-ene Of -14-yl] -carbamic acid methyl ester

Cyclopropanesulfonic acid [14-amino-18- (isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-ene in 2 mL DCM To a mixture of -4-carbonyl] -amide bishydrochloric acid (50 mg, 0.075 mmol) was added 44 μL (0.25 mmol) DIPEA and 9 mg (0.10 mmol) methyl chloroformate. The mixture was stirred at room temperature for 2 hours. It was then diluted with EtOAc and then washed with pH 4 buffer (2x) and brine (1x). The organic layer was dried (MgSO ₄ ) and then concentrated under reduced pressure to give the crude product. Flash chromatography (2% MeOH / DCM) gave 40 mg (82%) of the desired product as a white solid: LC-MS (Method A, retention time: 3.05 min), MS m / z 654 (M ⁺ +1).
Example 16 Compound 13, [4-cyclopropanesulfonylaminocarbonyl-18- (isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7- En-14-yl] -carbamic acid 2,2-dimethyl-propyl ester

Cyclopropanesulfonic acid [14-amino-18- (isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7- in 2 mL DCM To a mixture of en-4-carbonyl] -amide bishydrochloric acid (50 mg, 0.075 mmol) was added 44 μL (0.25 mmol) DIPEA and 15 mg (0.1 mmol) neopentyl chloroformate. The mixture was stirred at room temperature for 2 hours. It was then diluted with EtOAc and then washed with pH 4 buffer (2x) and brine (1x). The organic layer was dried (MgSO ₄ ) and then concentrated under reduced pressure to give the crude product. The obtained oil was dissolved in methanol and purified by preparative HPLC (YMC XTERRA, S5, 30x50 mm, gradient: 65% B to 100% B, 15 min, retention 2 min, flow rate 40 mL / min). The product was isolated (42 mg, 79%): LC-MS (Method A, retention time: 3.58 min), MS m / z 710 (M ⁺ +1).
General production method of chloroformate

This method was used to produce a chloroformate that is not commercially available. To a solution of 5.96 g (67.6 mmol) of commercial reagent (S) -3-hydroxytetrahydrofuran and pyridine (5.8 mL; 72 mmol) in THF (150 mL) cooled to 0 ° C. under argon, toluene (48 mL, 92.6 mmol). A 1.93M solution of phosgene in) was added over 10 minutes. The resulting solution was warmed at room temperature for 2 hours, the resulting solid was filtered, and then the mother liquor was carefully concentrated under reduced pressure at room temperature until the theoretical amount was obtained. The resulting residue was dissolved in 100 mL of THF to produce a 0.68M stock solution of 3 (S) -oxo-tetrahydrofuran chloroformate and stored in a freezer until use. In a similar manner, other commercial alcohols could be converted to the corresponding chloroformate 0.68M stock solution.
Example 17 Compound 14, [4-cyclopropanesulfonylaminocarbonyl-18- (isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7- En-14-yl] -carbamic acid tetrahydro-pyran-4-yl ester

Cyclopropanesulfonic acid [14-amino-18- (isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] in DCM prepared as described above. Nonadeca-7-en-4-carbonyl] -amide bishydrochloric acid (50 mg, 0.075 mmol, 44 μL (0.25 mmol) DIPEA, and 0.17 mL (0.1 mmol) tetrahydropyran chloroformate (tetrahydro-pyran-4-ol To give 28 mg (52%) of a white solid, preparative HPLC conditions: YMC XTERRA, S5, 19x100 mm, gradient: 50% B to 100% B, 10 min, retention 2 min. LC-MS (Method A, retention time: 3.13 min), MS m / z 724 (M ⁺ +1).
Example 18 Compound 15, [4-cyclopropanesulfonylaminocarbonyl-18- (isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7- En-14-yl] -carbamic acid tetrahydro-furan-3-yl ester

Cyclopropanesulfonic acid [14-amino-18- (isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] in DCM prepared as described above. Nonadeca-7-en-4-carbonyl] -amide bishydrochloric acid (50 mg, 0.075 mmol), 44 μL (0.25 mmol) DIPEA, and 0.10 mL (0.1 mmol) tetrahydro-furan-3-yl chloroformate (tetrahydro -Furan-3-ol was prepared by treating with phosgene) to give 28 mg (52%) of a white solid. Preparative HPLC conditions: YMC ODS-A, S5, 30x50mm, gradient: 50% B to 85% B, 8 min, retention 2 min, flow rate 45 ml / min. LC-MS (Method A, retention time: 3.05 min), MS m / z 710 (M ⁺ +1).
Example 19 Compound 16, [4-cyclopropanesulfonylaminocarbonyl-18- (isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7- En-14-yl] -carbamic acid isopropyl ester

Cyclopropanesulfonic acid [14-amino-18- (isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7 as described in the above method -En-4-carbonyl] -amide bishydrochloric acid (50 mg, 0.075 mmol), 44 μL (0.25 mmol) DIPEA, and 0.10 mL (0.1 mmol) 1M isopropyl chloroformate in toluene (Aldrich), 40 mg (78%) of white solid was obtained. Preparative HPLC conditions: YMC XTERRA, S5, 30x50 mm, gradient: 50% B to 100% B, 15 min, retention 2 min, flow rate 40 mL / min) NMR (300 MHz, CD ₃ OD) δ. LC-MS (Method A, retention Time: 3.32 min), MS m / z 682 (M ⁺ +1).
Example 20 (1S, 4R, 6S, 14S, 18R) -7-cis-14-tert-butoxycarbonylamino-18-hydroxy-2,15-dioxo-3,16 used in the preparation of compound 17 of Example 21 -Diazatricyclo [14.3.0.0 ^4,6 ] nonadeca-7-ene-4-carboxylic acid production

工程20A：1-(2(S)-tert-ブトキシカルボニルアミノ-ノナ-8-エノイル)-4(R)-ヒドロキシ-ピロリジン-2(S)-カルボン酸メチルエステルの製造

Step 20A : Preparation of 1- (2 (S) -tert-butoxycarbonylamino-nona-8-enoyl) -4 (R) -hydroxy-pyrrolidine-2 (S) -carboxylic acid methyl ester

A solution of 2 (S) -tert-butoxycarbonylamino-8-nonenoic acid (purchased from RSP A, mino Acids) (3.5 g, 12.9 mmol) in 200 mL DCM was sequentially added to 4 (R) -hydroxypyrrolidine-2 ( Treated with S) -carboxylic acid methyl ester / hydrochloric acid (2.15 g, 11.8 mmol), N-methylmorpholine (4.25 mL, 38.6 mmol), and HATU (5.37 g, 14.1 mmol). The reaction mixture was stirred at room temperature for 3 days under N ₂ and then concentrated under reduced pressure. The residue was partitioned between ethyl acetate and pH 4 buffer (biphthalate). The organic layer was washed with sat. Aq. NaHCO ₃ , dried (MgSO ₄ ) and then concentrated under reduced pressure to give the crude product. By flash chromatography (50% ethyl acetate / hexane to 100% ethyl acetate) 4.7 g (-100%) of 1- (2 (S) -tert-butoxycarbonylamino-nona-8-enoyl) -4 (R)- Hydroxy-pyrrolidine-2 (S) -carboxylic acid methyl ester was obtained as a colorless oil: ¹ H NMR (500 MHz, CD ₃ OD) δ 1.33-1.50 (m, 8H), 1.46 (s, 9H), 1.57 ( m, 1H), 1.72 (m, 1H) 2.08 (m, 2H), 2.28 (m, 1H), 3.72 (s, 3H,) 3.75-3.87 (m, 2H), 4.36 (m, 1H), 4.51 (bs, 1H), 4.57 (t, J = 8.2Hz, 1H), 4.95 (d, J = 10.4Hz, 1H), 5.01 (m, 1H), 5.83 (m, 1H). LC-MS (Method A , Retention time: 3.01 min), MS m / z 399 (M ⁺ +1).
Step 20B : 1-{[1- (2 (S) -tert-butoxycarbonylamino-nona-8-enoyl) -4 (R) -hydroxy-pyrrolidine-2 (S) carbonyl]-(1R) -amino} -2 (S) -Vinyl-cyclopropanecarboxylic acid ethyl ester production

1- (2 (S) -tert-butoxycarbonylamino-nona-8-enoyl) -4 (R) -hydroxy-pyrrolidine-2 (S) -carboxylic acid methyl ester (4.7 g, 11.8 mmol) in THF (80 mL ), Methanol (20 mL), and water (40 mL). Powdered lithium hydroxide (5.6 g, 233 mmol) was added. The pale yellow slurry was stirred under N ₂ at room temperature for 16 hours and then concentrated under reduced pressure. The residue was partitioned between ether and water. The ether phase was discarded and the aqueous phase was treated with 1N HCl until the pH was 4. The acidic solution was extracted with EtOAc (3x). The combined EtOAc extracts were dried (MgSO ₄ ) and then concentrated under reduced pressure to give 4.36 g (96%) of 1- (2 (S) -tert-butoxycarbonylamino-8-nonenoyl) -4 (R)- Hydroxy-pyrrolidine-2 (S) -carboxylic acid was obtained as a white solid. This acid was then dissolved in 150 mL of DMF, then (1R, 2S) -1-amino-2-vinylcyclopropanecarboxylic acid ethyl ester / hydrochloric acid (2.61 g, 13.6 mmol), N-methylmorpholine (2.5 mL 22.6 mmol), and HATU (5.2 g, 13.7 mmol). The reaction mixture was stirred at room temperature for 16 hours under N ₂ and then concentrated under reduced pressure. The residue was partitioned between ethyl acetate and pH 4 buffer (biphthalate). The organic layer was washed with sat. Aq. NaHCO ₃ , dried (MgSO ₄ ) and then concentrated under reduced pressure to give the crude product. By flash chromatography (60% -80% ethyl acetate / hexane) 6.0 g (98%) of 1-{[1- (2 (S) -tert-butoxycarbonylamino-non-8-enoyl) -4 (R) -Hydroxy-pyrrolidine-2 (S) carbonyl]-(1R) -amino} -2 (S) -vinyl-cyclopropanecarboxylic acid ethyl ester was obtained as a white solid: ¹ H NMR (500 MHz, CD ₃ OD) δ 1.25 (t, J = 7.2Hz, 3H), 1.33-1.80 (m, 10H), 1.46 (s, 9H), 2.09 (m, 3H), 2.25 (m, 2H), 3.76 (m, 2H), 4.14 (m, 2H), 4.27 (dd, J = 8.5, 5.2Hz, 1H), 4.50 (m, 2H), 4.94 (d, J = 10.1Hz, 1H), 5.01 (dd, J = 17.1, 1.8Hz, 1H), 5.11 (dd, J = 10.4, 1.8Hz, 1H), 5.30 (d, J = 15.6Hz, 1H), 5.80 (m, 2H), 8.57 (s, 1H). LC-MS (Method A, Retention time: 3.21 min), MS m / z 522 (M ⁺ +1).
Step 20C : (1S, 4R, 6S, 14S, 18R) -7-cis-14-tert-butoxycarbonylamino-
Preparation of 18-hydroxy-2,15-dioxo-3,16-diazatricyclo [14.3.0.0 ^4,6 ] nonadeca-7-ene-4-carboxylic acid ethyl ester

1-{[1- (2 (S) -tert-butoxycarbonyl-amino-nona-8-enoyl) -4 (R) -hydroxy-pyrrolidine-2 (S) carbonyl]-(in 2 L of methylene chloride A solution of 1R) -amino} -2 (S) -vinylcyclopropane-carboxylic acid ethyl ester (800 mg, 1.53 mmol) was flushed with N ₂ for 0.5 h. Next, tricyclohexylphosphine [1,3-bis (2,4,6-trimethyl-phenyl) -4,5-dihydroimidazol-2-ylidene] [benzylidene] -ruthenium (IV) dichloride (Strem) (64 mg, 0.075 mmol) was added and the mixture was then flushed with N ₂ for an additional 10 minutes. The pale orange homogeneous solution was refluxed for 2 hours to give a dark orange solution. The reaction mixture was cooled to room temperature and then concentrated under reduced pressure to give an orange oil. 460 mg (61%) of (1S, 4R, 6S, 14S, 18R) -7-cis-14-tert-butoxycarbonylamino-18-hydroxy-2,15-dioxo-3,16 by flash chromatography (ethyl acetate) -Diazatricyclo [14.3.0.0 ^4,6 ] -nonadeca-7-ene-4-carboxylic acid ethyl ester was obtained as a gray solid. ¹ H NMR (500 MHz, CDCl ₃ ) δ 1.19 (t, J = 7.2 Hz, 3H), 1.42 (s, 9H), 1.22-1.8 (m, 8H), 1.87 (m, 2H), 2.03-2.22 (m , 4H), 2.63 (m, 1H), 3.65 (m, 1H), 4.09 (m, 3H), 4.45 (m, 1H), 4.56 (s, 1H), 4.82 (m, 1H), 5.23 (m, 1H), 5.51 (s, 1H), 7.16 (s, 1H). LC-MS (Method A, retention time: 2.97 min), MS m / z 494 (M ⁺ +1).
Step 20D : (1S, 4R, 6S, 14S, 18R) -7-cis-14-tert-butoxycarbonylamino-18-hydroxy-2,15-dioxo-3,16-diazatricyclo [14.3.0.0 ^4,6 ] -Nonadeca-7-ene-4-carboxylic acid

(1S, 4R, 6S, 14S, 18R) -7-cis-14-tert-butoxycarbonylamino-18-hydroxy-2,15- in THF (4 mL), methanol (1 mL), and water (2 mL) Powdered lithium hydroxide (480 mg, 20 mmol) was added to a solution of dioxo-3,16-diazatricyclo [14.3.0.0 ^4,6 ] -nonadeca-7-ene-4-carboxylic acid ethyl ester (493 mg, 1.0 mmol), and lightly added. The yellow slurry was stirred at room temperature under N ₂ for 16 hours. The mixture was then concentrated under reduced pressure and the residue was partitioned between ether and water. The ether phase was discarded and the aqueous phase was treated with 1 N HCl until pH4. The acidic solution was extracted 3 times with EtOAc. The combined EtOAc extracts were dried (MgSO ₄ ) and then concentrated under reduced pressure to give 460 mg (98%) of Example 26, (1S, 4R, 6S, 14S, 18R) -7-cis-14-tert-butoxy Carbonylamino-18-hydroxy-2,15-dioxo-3,16-diazatricyclo [14.3.0.0 ^4,6 ] -nonadeca-7-ene-4-carboxylic acid was obtained as a gray solid. ¹ H NMR (500 MHz, CD ₃ OD) δ ppm 1.26 (t, J = 7.2 Hz, 3H), 1.35-1.52 (m, 15H), 1.57-1.68 (m, 3H), 1.79 (m, 1H), 2.04 (m, 1H), 2.16-2.41 (m, 3H), 3.80 (dd, J = 10.7, 4.3Hz, 1H), 3.88 (m, 1H), 4.38 (dd, J = 8.9, 3.1Hz, 1H), 4.55 (m, 2H), 5.39 (t, J = 9.8Hz, 1H), 5.58 (m, 1H). LC-MS (Method A, retention time: 2.64min), MS m / z 466 (M ⁺ +1 ).
Example 21 Compound 17B, [4-Cyclopropanesulfonylaminocarbonyl-18- (6-methoxy-isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] Nonadeca-7-en-14-yl] -carbamic acid tert-butyl ester

4-tert-Butoxycarbonylamino-18-hydroxy-2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-ene-4-carboxylic acid in DMSO (5 mL) To a mixture of 215 mg, 0.46 mmol) was added t-BuOK (125 mg, 1.11 mmol) and 1-chloro-6-methoxy-isoquinoline (110 mg, 0.56 mmol). The reaction was stirred at room temperature for 16 hours. The reaction mixture was then partitioned between ether (10 mL) and water (10 mL). The aqueous phase was acidified to pH 4 using 1N HCl. The resulting solution was extracted with EtOAc (3 × 20 mL). The combined EtOAc extracts were dried (MgSO ₄ ), filtered and then concentrated under reduced pressure to give a white solid. Flash chromatography (2% MeOH / CH ₂ Cl ₂ ) gave 140 mg (49%) of the carboxylic acid derivative as a white solid. LC-MS (Method B, retention time: 1.80 min), MS m / z 543 (M ⁺ +1). The solid (140 mg, 0.22 mmol) was treated with cyclopropylsulfonamide (35 mg, 0.28 mmol) as described in the general method above to give the crude product. Flash chromatography (2% MeOH / DCM) gave 90 mg of the desired product. Further purification by preparative HPLC (YMC Xterra, S5, 30 × 50 mm, 50% to 100% B, gradient 9 min, retention 1 min, flow rate 40 mL / min) gave 30 mg (19%) of product as a white powder: LC-MS (Method B, retention time: 1.86 min), MS m / z 726 (M ⁺ +1).
Example 22 1-{[4- (6-Methoxy-isoquinolin-1-yloxy) -pyrrolidine-2-carbonyl] -amino} -2-vinyl-cyclopropanecarboxylic acid ethyl ester (as bis-HCl) used in Example 24 )Manufacturing of

6-Methoxy-1-chloroquinoline (as described in Example 1, eg, in Steps 1a and 1b) was used to produce the product of Example 3 Step 2B except that 1-chloroquinoline was used. Prepared using the same synthetic procedure as
Example 23 Example 23 used in Example 24, Preparation of 2 (S) -tert-butoxycarbonylamino-3-pent-4-enylsulfanylpropionic acid

工程1：室温のメタノール (166mL)中のN-Boc-システインメチルエステル(3.36g、0.014mol)の溶液に、トリエチルアミン (10.8mL)および1-ブロモペンタ-4-エン (3.19g、21mmol、1.5当量)を加え、次いで得られた溶液を一夜室温で撹拌した。次に、混合物を減圧下で濃縮し、得られた残存混合物をフラッシュクロマトグラフィ(ヘキサン、酢酸エチル 勾配)で精製して1.76g(41%)の所望のチオエーテルを得た。¹H NMR (500MHz、CDCl₃) δ 1.43 (s、9H)、1.64 (m、2H)、2.11 (m、2H)、2.51 (m、2H)、2.95 (m、2H)、3.75 (s、3H)、4.51 (m、1H)、4.95-5.03 (m、2H)、5.34 (m、1H)、5.80 (1H、m)。LC-MS (方法B、ただし、勾配時間は3分間、流速は4ml/min、保持時間：2.29min)、MS m/z 304(M⁺+1)。
工程2：工程1のチオエーテル生成物 (9.51g、31.4mmol)を、水(200mL)およびTHF (200mL)中の1M LiOHの混合物に加え、得られた混合物を室温で一夜撹拌した。次に、反応混合物を1N塩酸を用いて酸性化し、得られた混合物を酢酸エチルで数回抽出した。抽出物を混合し、硫酸マグネシウムで乾燥し、次いで減圧下で濃縮して所望の酸を得、これをそのまま次の反応に用いた。
実施例24 化合物18B、[4-シクロプロパンスルホニルアミノカルボニル-18-(6-メトキシ-イソキノリン-1-イルオキシ)-2,15-ジオキソ-12-チア-3,16-ジアザ-トリシクロ[14.3.0.0^4,6]ノナデカ-7-エン-14-イル]-カルバミン酸tert-ブチルエステルの製造

Step 1 : To a solution of N-Boc-cysteine methyl ester (3.36 g, 0.014 mol) in methanol (166 mL) at room temperature, triethylamine (10.8 mL) and 1-bromopent-4-ene (3.19 g, 21 mmol, 1.5 eq) ) And then the resulting solution was stirred overnight at room temperature. The mixture was then concentrated under reduced pressure and the resulting residual mixture was purified by flash chromatography (hexane, ethyl acetate gradient) to give 1.76 g (41%) of the desired thioether. ¹ H NMR (500 MHz, CDCl ₃ ) δ 1.43 (s, 9H), 1.64 (m, 2H), 2.11 (m, 2H), 2.51 (m, 2H), 2.95 (m, 2H), 3.75 (s, 3H ), 4.51 (m, 1H), 4.95-5.03 (m, 2H), 5.34 (m, 1H), 5.80 (1H, m). LC-MS (Method B, where gradient time is 3 minutes, flow rate is 4 ml / min, retention time: 2.29 min), MS m / z 304 (M ⁺ +1).
Step 2 : The thioether product of Step 1 (9.51 g, 31.4 mmol) was added to a mixture of 1M LiOH in water (200 mL) and THF (200 mL) and the resulting mixture was stirred at room temperature overnight. The reaction mixture was then acidified with 1N hydrochloric acid and the resulting mixture was extracted several times with ethyl acetate. The extracts were combined, dried over magnesium sulfate, then concentrated under reduced pressure to give the desired acid, which was used as such for the next reaction.
Example 24 Compound 18B, [4-Cyclopropanesulfonylaminocarbonyl-18- (6-methoxy-isoquinolin-1-yloxy) -2,15-dioxo-12-thia-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] Preparation of nonadeca-7-en-14-yl] -carbamic acid tert-butyl ester

工程24A 1-{[1-(2-tert-ブトキシカルボニルアミノ-3-ペンタ-4-エニルスルファニル-プロピオニル)-4-(6-メトキシ-イソキノリン-1-イルオキシ)-ピロリジン-2-カルボニル]-アミノ}-2-ビニル-シクロプロパンカルボン酸エチルエステルの製造

Step 24A 1-{[1- (2-tert-Butoxycarbonylamino-3-pent-4-enylsulfanyl-propionyl) -4- (6-methoxy-isoquinolin-1-yloxy) -pyrrolidine-2-carbonyl]- Preparation of amino} -2-vinyl-cyclopropanecarboxylic acid ethyl ester

To a solution of the carboxylic acid of Example 23 (289 mg, 1.00 mmol, 1 eq) in DMF (9 mL), the dipeptide product of Example 22 (498 mg, 1.00 mmol, 1 eq), HATU (456 mg, 1.20 mmol, 1.2 eq), and N-methylmorpholine (385 μL, 3.50 mmol, 3.5 eq) were added. The solution was stirred overnight at room temperature, at which time LC / MS showed the formation of product and the disappearance of starting material 2. A pH 4 buffer solution was added and then the mixture was extracted three times with ethyl acetate. The organic layer was washed with brine, dried over magnesium sulfate and then concentrated under reduced pressure. Chromatography was performed (silica gel, ethyl acetate / hexane gradient) to give a quantitative amount of pure tripeptide 3.
¹ H NMR (400 MHz, CDCl ₃ ) δ 8.03 (d, J = 9.3 Hz, 1H), 7.89 (d, J = 5.9 Hz, 1H), 7.56 (br s, 1H), 7.15-7.09 (m, 2H) , 7.01 (d, J = 2.5Hz, 1H), 5.86-5.68 (m, 3H), 5.39-5.22 (m, 2H), 5.12-4.82 (m, 4H), 4.61 (app q, J = 6.8Hz, 1H), 4.24-4.04 (m, 3H), 3.91 (s, 3H), 3.02-2.90 (m, 1H), 2.842.69 (m, 2H), 2.66 (m, 3H), 2.24-2.08 (m, 2H), 1.88 (dd, J = 5.5, 8.0Hz, 1H), 1.68 (app quint, J = 7.5Hz, 2H), 1.56 (dd, J = 5.4, 9.5Hz, 1H), 1.42 (m, 1H) , 1.31 (s, 9H), 1.27-1.20 (m, 4H). [M / z] + H 697, ERRA 3.0X50mm S7 Retention time = 1.937 min, HPLC method 2 min gradient 0% B to 100% B, then 100% B for 1 minute (total operating time 3 minutes).
Step 24B 14-tert-Butoxycarbonylamino-18- (6-methoxy-isoquinolin-1-yloxy) -2,15-dioxo-12-thia-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca Of 7-ene-4-carboxylic acid ethyl ester

To a solution of the tripeptide product of step 24A (384.6 mg, 553 μmol, 1 eq) in dichloroethane (159 mL) was added tricyclohexylphosphine [1,3-bis (2,4,6-trimethylphenyl) -4,5 -Dihydroimidazol-2-ylidene [benzylidene] ruthenium (IV) dichloride (48.5 mg, 57 μmol, 0.10 eq) was added and refluxed for 16 hours. The reaction was concentrated under reduced pressure. Chromatography was performed (silica gel, ethyl acetate / hexane gradient) to give a pure macrocycle in 73% yield.
¹ H NMR (500 MHz, CDCl ₃ ) δ 8.04 (d, J = 9.2 Hz, 1H), 7.86 (d, J = 5.8 Hz, 1H), 7.59 (br s, 1H), 7.16-7.09 (m, 2H) , 7.00 (d, J = 2.4Hz, 1H), 5.78-5.71 (m, 1H), 5.64 (d, J = 7.6Hz, 1H), 5.58-5.50 (m, 1H), 5.36 (m, 1H), 5.03 (dd, J = 4.2, 8.3Hz, 1H), 4.81-4.75 (m, 1H), 4.27-4.14 (m, 2H), 3.98-3.93 (m, 1H), 3.91 (s, 3H), 3.13 3.02 (m, 2H), 3.00-2.94 (m, 1H), 2.69 (dt, J = 3.9, 11.2Hz, 1H), 2.52-2.42 (m, 2H), 2.36-2.26 (m, 2H), 2.16 2.08 (m, 1H), 1.96 (dd, J = 5.5, 8.2Hz, 1H), 1.86-1.76 (m, 1H), 1.68-1.58 (m, 2H), 1.37 (s, 9H), 1.29 (t, J = 7.2Hz, 1H), 1.27-1.21 (m, 1H).
[m / z] + H 669
XTERRA 3.0X50mm S7 Retention time = 1.797, HPLC method 2 min gradient 0% B to 100% B, then 100% B for 1 min (total run time 3 min).
Step 24C 18A, 14-tert-butoxycarbonylamino-18- (6-methoxy-isoquinolin-1-yloxy) -2,15-dioxo-12-thia-3,16-diaza-tricyclo [14.3.0.0 ^4,6 Production of nonadeca-7-ene-4-carboxylic acid

LiOH (97 mg, 4.04 mmol, 10 eq), water (0.45 mL), and MeOH (1.9 mL) in a solution of the macrocyclic ester product of step 24B (271 mg, 406 μmol, 1 eq) in THF (4.1 mL) Was added. The reaction was stirred for 16 hours, at which time LC / MS indicated that the hydrolysis was complete. A 1M solution of HCl in water was added and the resulting mixture was extracted three times with ethyl acetate. The organic extract was washed with brine, dried over magnesium sulfate and then concentrated under reduced pressure to give a quantitative amount of pure carboxylic acid.
¹ H NMR (400 MHz, DMSO-d ₆ ) δ 12.50 (br s, 1H), 8.59 (br s, 1H), 8.22 (d, J = 7.8 Hz, 1H), 8.14-8.07 (m, 1H), 7.46 (br s, 2H), 7.33-7.25 (m, 2H), 5.95 (br s, 1H), 5.79-5.56 (m, 2H), 4.66-4.57 (m, 1H), 4.50-4.37 (m, 2H) , 4.24-4.11 (m, 2H), 4.05 (s, 3H), 3.60-3.40 (m, 2H), 3.09-2.97 (m, 1H), 2.91.2.81 (m, 1H), 2.65-2.58 (m, 1H), 2.56-2.44 (m, 2H), 2.44-2.33 (m, 1H), 1.84-1.73 (m, 1H), 1.72-1.57 (m, 2H), 1.34 (s, 9H), 1.08-0.95 ( m, 1H).
[m / z] + H 641
XTERRA 3.0X50mm S7 Retention time = 1.673, HPLC method 2 min gradient 0% B-100% B, then 100% B for 1 min (total run time 3 min).
Step 24D Compound 18B, [4-Cyclopropanesulfonylaminocarbonyl-18- (6-methoxy-isoquinolin-1-yloxy) -2,15-dioxo-12-thia-3,16-diaza-tricyclo [14.3.0.0 ^{4 , 6} ] Preparation of nonadeca-7-en-14-yl] -carbamic acid tert-butyl ester

To a solution of the carboxylic acid product of Step 24C (236.3 mg, 369 μmol, 1 eq) in THF (5.2 mL) was added carbonyldiimidazole (143 mg, 738 μmol, 2 eq) and then refluxed for 2.5 hours. The solution was cooled to room temperature at which time cyclopropylsulfonamide (158 mg, 1.1 mmol, 3 eq) and DBU (127 μL, 849 μmol, 2.3 eq) were added. The reaction was stirred at room temperature for 16 hours. A 1M solution of HCl in water was added and the product was then extracted three times with methylene chloride. The organic layer was washed with brine, dried over magnesium sulfate and then concentrated under reduced pressure. Chromatography was performed (silica gel, ethyl acetate / hexane gradient) to give pure sulfonamide compound 18 in 78% yield.
¹ H NMR (400 MHz, CD ₃ OD) δ 8.10 (d, J = 9.3 Hz, 1H), 7.89-7.83 (m, 2H), 7.21 (d, J = 5.9 Hz, 1H), 7.13 (br s, 1H ), 7.05 (d, J = 9.1Hz, 1H), 5.81 (br s, 1H), 5.68 (app q, J = 8.9Hz, 1H), 5.16 (app t, J = 9.7Hz, 1H), 4.59- 4.52 (m, 2H), 4.40-4.32 (m, 1H), 4.12-4.04 (m, 1H), 3.89 (s, 3H), 2.98-2.78 (m, 3H), 2.73-2.41 (m, 6H), 2.02-1.98 (m, 1H), 1.75 (dd, J = 5.5, 8.2Hz, 1H), 1.71-1.53 (m, 3H), 1.32-1.20 (m, 1H), 1.16 (s, 9H), 1.14 0.95 (m, 2H).
[m / z] + H 744
XTERRA 3.0X50mm, S7 retention time = 1.700, HPLC method 2 min gradient 0% B-100% B, then 100% B for 1 min (total run time 3 min).
Example 25 Compound 19, 4-cyclopropanesulfonylaminocarbonyl-18- (6-methoxy-isoquinolin-1-yloxy) -2,12,12,15-tetraoxo-12-thia-3,16-diaza-tricyclo [ 14.3.0.0 ^4,6 ] Production of nonadeca-7-en-14-yl] -carbamic acid tert-butyl ester

To a solution of compound 18 (84.7 mg, 114 μmol) in MeOH (1.4 mL) and water (460 μL) at 0 ° C. was added oxone (210.6 mg, 343 μmol, 3.0 eq). The mixture was stirred at 0 ° C. for 20 minutes and then at ambient temperature for 3 hours. Then water and methylene chloride were added and the layers were separated. The aqueous layer was extracted twice more with methylene chloride. The combined organic layer was dried over magnesium sulfate and then concentrated under reduced pressure. The product was purified by prep TLC (ethyl acetate) to give the sulfone (37.2 mg, 42% (yield)).
¹ H NMR (CD ₃ OD) δ 8.11 (d, J = 9.5Hz, 1H), 7.88 (d, J = 6.1Hz, 1H), 7.23 (d, J = 5.8Hz, 1H), 7.20-7.15 (m , 1H), 7.11 (dd, J = 1.8, 8.9Hz, 1H), 5.87-5.81 (m, 1H), 5.61-5.47 (m, 2H), 4.96-4.90 (m, 1H), 4.67 (t, J = 7.8Hz, 1H), 4.46-4.39 (m, 1H), 4.27-4.20 (m, 1H), 3.92 (s, 3H), 3.84-3.72 (m, 1H), 2.94.2.84 (m, 2H), 2.67-2.58 (m, 2H), 2.47-2.37 (m, 1H), 2.37-2.25 (m, 2H), 1.85-1.73 (m, 2H), 1.72-1.64 (m, 1H), 1.49-1.38 (m , 1H), 1.20 (s, 9H), 1.16-1.02 (m, 3H), 1.00-0.82 (m, 3H).
Example 26 Preparation of Compound 27 and Compound 28

工程26A：イソプロピルピロリジン-5-オン-2(S)-カルボキシレートの製造

Step 26A : Preparation of isopropylpyrrolidin-5-one-2 (S) -carboxylate

A solution of L-pyroglutamic acid (Aldrich, 25.0 g, 195 mmol) and para-toluenesulfonic acid monohydrate (3.71 g, 19.5 mmol) was added to the Dean-Stark trap variation (condensate 4x molecular sieve packed Soxhlet extract. Was refluxed in isopropanol (40 mL) under nitrogen for 6 hours. After cooling to room temperature, the reaction was diluted with ether, washed with saturated aqueous sodium bicarbonate, then saturated aqueous NaCl, dried (MgSO ₄ ), and evaporated to give a colorless syrup. It crystallized as soon as it hardened. The crystal residue was triturated in hexane to give 31.9 g (96%) of isopropylpyrrolidin-5-one-2 (S) -carboxylate as a white prism: ¹ H NMR (300 MHz, chloroform-D) δ 6.35 (br s, 1H), 5.04 (sept.1H, J = 6.2Hz), 4.18 (dd, 1H, J = 8.4, 5.3Hz), 2.51-2.28 (m, 3H), 2.27-2.12 (m, 1H ), 1.24 (d, 6H, J = 6.2 Hz). LCMS m / z 172 (M + H) ⁺ .
Step 26B : Preparation of isopropyl 1- (tert-butoxycarbonyl) -pyrrolidin-5-one-2 (S) -carboxylate

Isopropylpyrrolidin-5-one-2 (S) -carboxylate (product of Step 26A, 31.9 g, 188 mmol), di-tert-butyl dicarbonate (48.6 g, 225 mmol) and DMAP (in 300 mL) A solution of 2.30 g, 8.8 mmol) was stirred at room temperature under N ₂ for 30 minutes. The reaction is evaporated to about 100 mL, diluted with ether, washed with 1N HCl, then saturated aqueous NaCl, dried (MgSO ₄ ), then evaporated to isopropyl 1- (tert-butoxycarbonyl) pyrrolidin-5-one -2 (S) carboxylate was obtained as a pale yellow oil. 50.1 g (99%): ¹ H NMR (300 MHz, chloroform-D) δ 5.06 (sept. 1H, J = 6.2 Hz), 4.53 (dd, 1H, J = 9.5, 2.9 Hz), 2.66-2.40 (m, 2H), 2.36-2.22 (m, 1H), 2.03-1.93 (m, 1H), 1.47 (s, 9H), 1.26 (d, 3H, J = 6.2Hz), 1.24 (d, 3H, J = 6.2 Hz) ). LCMS m / z 272 (M + H) ⁺ .
Step 26C : Preparation of isopropyl 2 (S)-(tert-butoxycarbonylamino) -5-hydroxypentanoate

To a solution of isopropyl 1- (tert-butoxycarbonyl) pyrrolidin-5-one-2 (S) -carboxylate (product of Step 26B, 49.5 g, 183 mmol) in methanol (300 mL) was added sodium borohydride (10.0 g, 263 mmol) was added in ˜1 g portions over 1.5 hours. The reaction was stirred for another 10 minutes under nitrogen. It was diluted with water and extracted with ether, the organic fractions were combined, washed with saturated aqueous NaCl, dried (MgSO ₄ ) then evaporated to give a pale yellow oil. Flash chromatography (silica gel, 20-30% ethyl acetate / hexane) gave 31.8 g (64%) of isopropyl 2 (S)-(tert-butoxycarbonylamino) -5-hydroxypentanoate as a colorless syrup. : ¹ H NMR (300 MHz, chloroform-D) δ 5.16 (br d, 1 H, J = 7.3 Hz), 5.03 (sept., 1 H, J = 6.2 Hz), 4.28 (br d, 1 H, J = 6.2 Hz) , 3.67 (br dd, J = 10.2, 5.5Hz), 1.94-1.79 (m, 2H), 1.76-1.67 (m, 1H), 1.66-1.56 (m, 2H), 1.43 (s, 9H), 1.25 ( d, 3H, J = 6.2 Hz), 1.23 (d, 3H, J = 6.2 Hz). LCMS m / z 276 (M + H) ⁺ .
Step 26D : Preparation of isopropyl-5-allyloxy-2 (S)-(tert-butoxycarbonylamino) pentanoate

Isopropyl 2 (S)-(tert-butoxycarbonylamino) -5-hydroxypentanoate (product of Step 26C, 17.6 g, 63.9 mmol), allyl methyl carbonate (24.0 ml, 213 mmol) in THF (150 mL) , Pd ₂ (dba) ₃ (1.62 g, 1.78 mmol) and BINAP (4.42 g, 7.10 mmol) were refluxed under nitrogen for 3 hours. After cooling to room temperature, the reaction was diluted with ether, filtered through celite, then evaporated to give a dark brown syrup. The residue was subjected to flash chromatography (silica gel, 30% ether / hexane) to give isopropyl 5-allyloxy-2 (S)-(tert-butoxycarbonylamino) pentanoate as a viscous colorless oil. 16.3 g (81%): ¹ H NMR (300 MHz, chloroform-D) δ 5.88 (ddt, 1H, 17.4, 10.4, 5.5), 5.28 (m, 1H), 5.22-5.11 (m, 1H), 5.02 (sept ., 1H, J = 6.2Hz), 4.21 (br t, 1H, J = 6.7Hz), 3.94 (dt, 2H, J = 5.9, 1.5Hz), 3.42 (t, 2H, J = 5.9Hz), 1.90 -1.82 (m, 1H), 1.75-1.57 (m, 3H), 1.42 (s, 9H), 1.21 (d, 3H, J = 6.2Hz), 1.19 (d, 3H, J = 6.2 Hz). LCMS m / z 316 (M + H) ⁺ .
Step 26E : Preparation of 5-allyloxy-2 (S)-(tert-butoxycarbonylamino) pentanoic acid

Isopropyl 5-allyloxy-2 (S)-(tert-butoxycarbonylamino) pentanoate (product of Step 26D, 16.1 g, 51.1 mmol) and lithium hydroxide hydrate (THF / water (100 mL / 20 mL)) 4.19 g, 102 mmol) was stirred at room temperature under nitrogen for 16 hours. The reaction was diluted with water, washed with ether, adjusted to pH of the aqueous fraction to ~ 4, extracted with ether, the organic fractions were combined, washed with saturated NaCl, dried (MgSO _4), Evaporation then gave 5-allyloxy-2 (S)-(tert-butoxycarbonylamino) pentanoic acid as a pale yellow syrup: ¹ H NMR (300 MHz, chloroform-D) δ 5.89 (ddt, 1H, J = 17.4, 10.4, 5.5), 5.25 (dd, 1H, J = 17.4, 1.6Hz), 5.17 (dd, 1H, J = 10.4, 1.6Hz), 4.30 (br d, 1H, J = 6.2), 3.96 ( (dt, 2H, J = 5.9, 1.5Hz), 3.46 (t, 2H, J = 5.9Hz), 1.96-1.86 (m, 1H), 1.85-1.77 (m, 1H), 1.75-1.64 (m, 2H) 1.43 (s, 9 H). LCMS m / z 274 (M + H) ⁺ .
Step 26F : 1-{[1- (5-allyloxy-2-tert-butoxycarbonylamino-pentanoyl) -4- (6-methoxy-isoquinolin-1-yloxy) -pyrrolidine-2-carbonyl] -amino} -2 -Vinyl-cyclopropanecarboxylic acid ethyl ester production

1-{[4- (6-Methoxy-isoquinolin-1-yloxy) -pyrrolidine-2-carbonyl] -amino} -2-vinyl-cyclopropanecarboxylic acid ethyl ester dihydrochloride (Example 22) in 75 mL of DMF ) (3.60 g, 7.25 mmol) and 5-allyloxy-2-tert-butoxycarbonylamino-pentanoic acid (product of Step 26E, 1.98 g, 7.25 mmol) was added HATU (3.31 g, 8.7 mmol) and N -Treated with methylmorpholine (2.79 ml, 25.4 mmol). The reaction mixture was stirred at room temperature under N ₂ for 4 hours and then quenched with pH 4 buffer (biphthalate). The resulting mixture was extracted with EtOAc. The organic layer was washed with sat. Aq. NaCl, dried (MgSO ₄ ) and then concentrated under reduced pressure to give the crude product. By flash chromatography (silica gel, 10-60% ethyl acetate / hexane) 3.91 g (-79%) of 1-{[1- (5-allyloxy-2-tert-butoxycarbonylamino-pentanoyl) -4- (6- Methoxy-isoquinolin-1-yloxy) -pyrrolidine-2-carbonyl] -amino} -2-vinyl-cyclopropanecarboxylic acid ethyl ester was obtained as a yellow disintegrating foam: ¹ H NMR (500 MHz, methanol-D4) δ 1.05-1.16 (m, 1H), 1.32 (s, 9H), 1.37-1.50 (m, 3H), 1.51-1.67 (m, 3H), 1.68-1.84 (m, 2H), 2.17-2.30 (m, 1H), 2.32-2.50 (m, 1H), 2.58-2.73 (m, 1H), 2.80 (s, 1H), 3.33-3.52 (m, 2H), 3.86-3.95 (m, 5H), 4.00-4.06 ( m, 1H), 4.07-4.22 (m, 3H), 4.23-4.38 (m, 1H), 4.58-4.67 (m, 1H), 5.04-5.17 (m, 2H), 5.18-5.34 (m, 2H), 5.65-5.94 (m, 3H), 7.07-7.28 (m, 3H), 7.82-7.92 (m, 1H), 7.96-8.13 (m, 1H). LC-MS (Method B, retention time: 1.75 min), MS m / z 681 (M ⁺ +1).
Step 26G : 14-tert-butoxycarbonylamino-18- (6-methoxy-isoquinolin-1-yloxy) -2,15-dioxo-10-oxa-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] Preparation of nonadeca-7-ene-4-carboxylic acid ethyl ester

1-{[1- (5-Allyloxy-2-tert-butoxycarbonylamino-pentanoyl) -4- (6-methoxy-isoquinolin-1-yloxy) -pyrrolidine-2-carbonyl] -amino in 800 mL of benzene A solution of} -2-vinyl-cyclopropanecarboxylic acid ethyl ester (product of Step 26F, 2.0 g, 2.94 mmol) was treated with Grubb second generation catalyst (375 mg, 0.44 mmol). The reaction mixture was heated to 50 ° C., stirred for 2 h under N ₂ and then concentrated under reduced pressure. Flash chromatography (silica gel, 25-100% ethyl acetate / hexane) followed by treatment with activated charcoal and 677.6 mg (~ 35%) of 14-tert-butoxycarbonylamino-18- (6-methoxy-isoquinolin-1-yloxy) ) -2,15-dioxo-10-oxa-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-ene-4-carboxylic acid ethyl ester was obtained as a brown disintegrating foam: ¹ H NMR (500 MHz, methanol-D4) δ 1.06-1.30 (m, 12H), 1.51-2.03 (m, 6H), 2.41-2.70 (m, 3H), 3.39-3.50 (m, 1H), 3.51-3.61 (m, 1H), 3.77-3.87 (m, 1H), 3.92 (s, 3H), 4.02-4.18 (m, 3H), 4.24 (d, J = 6.41Hz, 1H), 4.36-4.48 (m, 1H ), 4.55 (d, J = 11.29Hz, 1H), 4.65 (t, J = 8.39Hz, 1H), 5.55-5.72 (m, 2H), 5.85 (s, 1H), 7.08 (d, J = 8.85Hz , 1H), 7.17 (s, 1H), 7.24 (d, J = 5.80Hz, 1H), 7.89 (d, J = 5.80Hz, 1H), 8.16 (d, J = 9.16Hz, 1H). LC-MS (Method B, retention time: 1.58 min), MS m / z 653 (M ⁺ +1).
Step 26H : Compound 27, 14-tert-butoxycarbonylamino-18- (6-methoxy-isoquinolin-1-yloxy) -2,15-dioxo-10-oxa-3,16-diaza-tricyclo [14.3.0.0 ^{4 , 6} ] Production of nonadeca-7-ene-4-carboxylic acid

14-tert-Butoxycarbonylamino-18- (6-methoxy-isoquinolin-1-yloxy) -2,15-dioxo-10-oxa-3,16-diaza-tricyclo [14.3.0.0 in 25.7 mL of THF ^4,6 ] A solution of nonadeca-7-ene-4-carboxylic acid ethyl ester (product of Step 26G, 667 mg, 1.02 mmol) was added to 2.9 mL water, 11.4 mL methanol, and powdered lithium hydroxide (489 mg, 20.4 mmol) was added. The reaction mixture was stirred at room temperature for 6 hours and then concentrated under reduced pressure. The residue was partitioned between ethyl acetate and 20.4 mL of 1 N aq. HCl. A pH 4 buffer (biphthalate) was added to the resulting solution. The organic layer was washed with water, sat. Aq. NaCl, dried (MgSO ₄ ) and then concentrated under reduced pressure to give the crude product. 446 mg (~ 70%) 14-tert-butoxycarbonylamino-18- (6-methoxy-isoquinolin-1-yloxy) -2,15-dioxo-10 by flash chromatography (silica gel, 0-10% methanol / chloroform) -Oxa-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-ene-4-carboxylic acid was obtained as a slightly pink disintegrating foam: ¹ H NMR (500 MHz, methanol -D4) δ 0.99-1.16 (m, 1H), 1.22 (s, 9H), 1.50-2.04 (m, 6H), 2.40-2.73 (m, 3H), 3.41-3.50 (m, 1H), 3.51-3.62 (m, 1H), 3.73-3.86 (m, 1H), 3.90 (s, 3H), 4.03 (d, J = 10.52Hz, 1H), 4.21 (d, J = 8.31Hz, 1H), 4.42-4.72 ( m, 3H), 5.65 (s, 2H), 5.82 (s, 1H), 7.06 (d, J = 9.05Hz, 1H), 7.15 (s, 1H), 7.22 (d, J = 5.87Hz, 1H), 7.87 (m, J = 9.29 Hz, 2H), 8.14 (d, J = 8.56 Hz, 1H). LC-MS (Method B, retention time: 1.46 min), MS m / z 625 (M ⁺ +1).
Step 26I : Compound 28, [4-Cyclopropanesulfonylaminocarbonyl-18- (6-methoxy-isoquinolin-1-yloxy) -2,15-dioxo-10-oxa-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] Preparation of nonadeca-7-en-14-yl] -carbamic acid tert-butyl ester

14-tert-butoxycarbonylamino-18- (6-methoxy-isoquinolin-1-yloxy) -2,15-dioxo-10-oxa-3,16-diaza-tricyclo [14.3.0.0 ^{4 in} 5 mL of THF ^{, 6]} 7-ene-4-carboxylic acid (the product of step 26H, 146.5mg, 0.235mmol) and carbonyl diimidazole (76 mg, 0.47 mmol) solution under N ₂ and stirred for 2 hours at reflux temperature And then brought to room temperature. To the reaction mixture was added cyclopropanesulfonic acid amide (86 mg, 0.71 mmol) and 1,8-diazabicyclo [5.4.0] undec-7-ene (81 μl, 0.54 mmol). The reaction mixture was stirred under N ₂ at room temperature for 4 hours and then quenched with pH 4 buffer (biphthalate). The resulting mixture was extracted with EtOAc. The organic layer was washed with sat. Aq. NaCl, dried (MgSO ₄ ) and then concentrated under reduced pressure to give the crude product. 111.7 mg (~ 65%) of (4-cyclopropanesulfonylaminocarbonyl-18- (6-methoxy-isoquinolin-1-yloxy) -2,15-dioxo by flash chromatography (silica gel, 0-8% methanol / chloroform) -10-Oxa-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-en-14-yl] -carbamic acid tert-butyl ester (compound 27) was obtained as a white powder: ¹ H NMR (500MHz, methanol-D4) δ 0.96-1.17 (m, 6H), 1.20 (s, 9H), 1.24-1.32 (m, 1H), 1.49-1.64 (m, 2H), 1.65-1.79 (m, 2H ), 1.79-2.06 (m, 2H), 2.45-2.60 (m, 1H), 2.62-2.76 (m, 2H), 2.6-2.98 (m, 1H), 3.47-3.61 (m, 2H), 3.62-3.73 (m, 1H), 3.92 (s, 3H), 4.00 (d, J = 10.38Hz, 1H), 4.18 (d, J = 10.99Hz, 1H), 4.55-4.72 (m, 3H), 5.44 (t, J = 9.92Hz, 1H), 5.69-5.78 (m, 1H), 5.86 (s, 1H), 7.07 (d, J = 7.02Hz, 1H), 7.17 (s, 1H), 7.24 (d, J = 5.80 Hz, 1H), 7.85-7.95 (m, 2H), 8.18 (d, J = 8.85Hz, 1H). LC-MS (Method B, retention time: 1.52 min), MS m / z 728 (M ⁺ +1).
Production of Compound 61

Compound 61 was produced using the following method as shown in the above reaction formula.
Production of Example 28

Example 28 was prepared by adding a DMF solution of N-trityl protected threonine to a DMF solution of sodium hydride cooled to -15 ° C. The reaction mixture was stirred at −15 ° C. for 30 minutes, then 5-bromo-1-pentene was added and the resulting mixture was warmed to −5 ° C. The reaction mixture was maintained at −5 ° C. for 3 days, then the reaction was quenched by addition of 1N aqueous HCl and then worked up using the standard extraction method described above. Example 28 was obtained in pure form by standard chromatographic methods.
Production of Example 29

Example 29 was prepared by coupling Example 28 with Example 22 as shown in the reaction scheme, using the structurally related compound and the general method described above for preparing the example.
Production of Example 30

Example 30 was prepared from Example 29 using the structurally related compounds and the general method described above for preparing the examples.
Production of Example 31

Example 31 was prepared from Example 30 as shown in the reaction scheme. In this case, 100 mg of Example 30 was dissolved in 2 mL of DCM and the resulting solution was treated with 82 μL of a 4: 1 solution of water and trifluoroacetic acid. The resulting mixture was stirred at room temperature for several hours, then the reaction mixture was concentrated and the resulting crude product mixture was purified by chromatography to give Example 31.
Production of Compound 61

Compound 61 was prepared from Example 31 using the methods described above for the preparation of the related compounds and examples as shown in the reaction scheme.
Preparation of Compound 62, Compound 66, and Compound 70

Compound 62 was prepared from compound 61 using the methods described above for the preparation of the related compounds and examples as shown in the reaction scheme.
Production of compounds 63, 64, 65, compounds 67, 68, 69, compounds 71, 72, 73

Compounds 63, 64, 65, 67, 68, 69, 71, 72 and 73 were prepared using the methods described above for the preparation of related compounds and examples, as shown in the above reaction scheme.
Example 36 : Preparation of compound 79 and compound 80

工程36A：N-t-ブトキシカルボニル-3-(4-ペンテニルチオ)-L-バリン、メチルエステルの製造

Step 36A : Preparation of Nt-butoxycarbonyl-3- (4-pentenylthio) -L-valine, methyl ester

  To a solution of 7.12 g (48 mmol, 1.0 eq) L-penicillamine in 100 mL 1,4-dioxane and 25 mL water at room temperature, add 9.60 mL (96 mmol, 2.0 eq) 10 N aqueous sodium hydroxide solution, then 12.00 mL (101 mmol, 2.1 eq) of 5-bromo-1-pentene was added dropwise over several minutes. The resulting mixture was stirred at room temperature for 68 hours. At this point, 12.50 g (57 mmol, 1.2 eq) of di-tert-butyl dicarbonate was added and the mixture was then stirred at room temperature for another 6 hours. The mixture was concentrated under reduced pressure and the residue was dissolved in water. The aqueous mixture was washed with diethyl ether, adjusted to pH 3 with 1N hydrochloric acid and then extracted with ethyl acetate. The combined extract was washed with brine, dried over anhydrous magnesium sulfate, filtered and then concentrated under reduced pressure.

The crude product (12.20 g) was dissolved in 120 mL anhydrous dimethyl sulfoxide. To this solution was added 10.50 g (76 mmol) of potassium carbonate and 4.70 mL (76 mmol) of iodomethane, and the resulting mixture was stirred at room temperature for 24 hours. The reaction mixture was diluted with water and then extracted with ethyl acetate. The combined extract was washed with water (2X) and brine, dried over anhydrous sodium sulfate, filtered and then concentrated under reduced pressure. Silica gel column chromatography (elution: 2-10% ethyl acetate / hexane) gave 8.54 g of N-tert-butoxycarbonyl-3- (4-pentenylthio) -L-valine, methyl ester as a colorless oil. NMR (300 MHz, CDCl ₃ ): δ 5.76 (d d t, 1H, J = 17.2, 10.3, 6.6 Hz), 5.35 (br d, 1H, J = 9.0 Hz), 5.05-4.94 (m, 2H) , 4.27 (br d, 1H, J = 9.0Hz), 3.73 (s, 3H), 2.52 (m, 2H), 2.13 (quart., 2H, J = 7.3Hz), 1.61 (quint., 2H, J = 7.3Hz), 1.43 (s, 9H), 1.35 (s, 3H), 1.33 (s, 3H).
Step 36B : Production of N-tert-butoxycarbonyl-3- (4-pentenylthio) -L-valine

A solution of 8.52 g (25.7 mmol) N-tert-butoxycarbonyl-3- (4-pentenylthio) -L-valine, methyl ester in 200 mL tetrahydrofuran at room temperature in 1.10 g (26.2 mmol) in 50 mL water. A solution of lithium hydroxide monohydrate was added. The resulting mixture was stirred at room temperature for 65 hours. Next, 28 mL of 1.00N hydrochloric acid was added to the reaction mixture. The mixture was diluted with diethyl ether, washed with water (3X) and brine, dried over anhydrous sodium sulfate, filtered and then concentrated under reduced pressure to 8.10 g of N-tert-butoxycarbonyl-3- (4- Pentenylthio) -L-valine was obtained as a colorless oil. NMR (300 MHz, CDCl ₃ ): δ 5.75 (d d t, 1H, J = 17.2, 10.3, 6.6 Hz), 5.40 (br s, 1H), 5.05-4.94 (m, 2H), 4.28 (br s , 1H), 2.56 (m, 2H), 2.13 (quart., 2H, J = 7.3Hz), 1.63 (quint., 2H, J = 7.3Hz), 1.44 (s, 9H), 1.39 (s, 3H) 1.37 (s, 3 H).
Step 36C : (1S, 4R, 6S, 14R, 18R) -7-cis-14-tert-butoxycarbonylamino-13,13-dimethyl-18- (6-methoxy-isoquinolin-1-yloxy) -2,15 Of 2-dioxo-12-thia-3,16-diazatricyclo [14.3.0.0 ^4,6 ] nonadeca-7-ene-4-carboxylic acid (Compound 79)

This compound was prepared from N-tert-butoxycarbonyl-3- (4-pentenylthio) -L-valine using the method described in Example 24. ESI-mass spectrum: m / e: 669 (M + H) ⁺ , 667 (MH) ⁻ ; high-resolution mass spectrum: theoretical value for C ₃₄ H ₄₅ N ₄ O ₈ S: 669.2958, actual value: 669.2975; NMR ( 500MHz, CD ₃ OD) δ 8.06 (d, 1H, J = 8.8Hz), 7.90 (d, 1H, J = 5.8Hz), 7.24 (d, 1H, J = 5.8Hz), 7.16 (s, 1H), 7.09 (d, 1H, J = 8.8Hz), 6.69 (d, 1H, J = 8.5Hz), 5.80 (s, 1H), 5.72 (quart., 1H, J = 8.5Hz), 5.46 (t, 1H, J = 8.5Hz), 4.71 (t, 1H, J = 8.5Hz), 4.64 (d, 1H, J = 9.1Hz), 4.45 (d, 1H, J = 11.6Hz), 4.22 (m, 1H), 3.91 (s, 3H), 2.78 (m, 1H), 2.74-2.68 (m, 2H), 2.56 (m, 1H), 2.36-2.28 (m, 2H), 2.21 (m, 1H), 1.75 (m, 1H ), 1.72-1.64 (m, 2H), 1.60 (m, 1H), 1.45 (s, 3H), 1.39 (s, 3H), 1.21 (s, 9H).
Step 36D : (1S, 4R, 6S, 14R, 18R)-[7-cis-4-cyclopropanesulfonyl-aminocarbonyl-13,13-dimethyl-18- (6-methoxyisoquinolin-1-yloxy) -2, Preparation of 15-dioxo-12-thia-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-en-14-yl] carbamic acid, tert-butyl ester (compound 80)

This compound was prepared from compound 79 using the method described in Example 24D. ESI-mass spectrum: m / e: 772 (M + H) ⁺ , 770 (MH) ⁻ ; high-resolution mass spectrum: theoretical value for C ₃₇ H ₅₀ N ₅ O ₉ S ₂ : 772.3050, actual value: 772.3060; NMR (500MHz, CD ₃ OD) δ 8.06 (d, 1H, J = 8.8Hz), 7.90 (d, 1H, J = 5.8Hz), 7.24 (d, 1H, J = 5.8Hz), 7.17 (s 1H), 7.08 (d, 1H, J = 8.8Hz), 6.62 (d, 1H, J = 8.5Hz), 5.83 (s, 1H), 5.74 (quart., 1H, J = 8.5Hz), 5.26 (t, 1H, J = 8.5Hz), 4.67 (m, 1H), 4.46 (m, 2H), 4.19 (m, 1H), 3.92 (s, 3H), 2.93 (m, 1H), 2.742.67 (m, 3H), 2.58-2.48 (m, 3H), 2.11 (m, 1H), 1.82-1.75 (m, 2H), 1.61 (m, 1H), 1.54 (m, 1H), 1.45 (s, 3H), 1.37 (s, 3H), 1.27 (m, 1H), 1.18 (s, 9H), 1.09 (m, 1H), 1.07-0.99 (m, 2H).
Example 37 : Preparation of compound 84 and compound 85

工程37A：N-tert-ブトキシカルボニル-O-(4-ペンテニル)-L-セリン、メチルエステルの製造

Step 37A : Production of N-tert-butoxycarbonyl-O- (4-pentenyl) -L-serine, methyl ester

  To a solution of 10.26 g (50 mmol, 1.0 eq) N-tert-butoxycarbonyl-L-serine in 500 mL anhydrous dimethyl sulfoxide, 2.00 g (50 mmol, 1.0 eq) 60% sodium hydride in mineral oil at room temperature Was added. The mixture was stirred at room temperature for 0.5 hour until gas evolution ceased. To the resulting solution was added 6.00 mL (50 mmol, 1.0 eq) of 5-bromo-1-pentene, immediately followed by another 2.00 g (50 mmol, 1.0 eq) of 60% sodium hydride in mineral oil. The reaction mixture was then stirred at room temperature for 16 hours. The mixture was diluted with 2000 mL water, adjusted to pH 3-4 with the addition of 50 mL 1.00N hydrochloric acid, and then extracted with ethyl acetate. The organic layer was washed with water (2X) and brine, dried over anhydrous sodium sulfate, filtered and then concentrated under reduced pressure. The material obtained to remove the remaining mineral oil was dissolved in dilute aqueous sodium hydroxide solution. The aqueous solution was washed with hexane, then adjusted to pH 4 with hydrochloric acid and then extracted with ethyl acetate. The extract was washed with water (2X) and brine, dried over anhydrous sodium sulfate, filtered and then concentrated under reduced pressure.

The crude product (7.70 g) was dissolved in 100 mL anhydrous dimethyl sulfoxide. To this solution was added 7.80 g (56 mmol) of potassium carbonate and 3.50 mL (56 mmol) of iodomethane and the resulting mixture was stirred at room temperature for 24 hours. The reaction mixture was diluted with water and extracted with ethyl acetate. The combined extract was washed with water (2X) and brine, dried over anhydrous sodium sulfate, filtered and then concentrated under reduced pressure. Silica gel column chromatography (elution: 2-10% ethyl acetate / hexane) gave 6.70 g of N-tert-butoxycarbonyl-O- (4-pentenyl) -L-serine, methyl ester as a colorless oil. NMR (300 MHz, CDCl ₃ ): δ 5.78 (d d t, 1H, J = 17.2, 10.2, 6.6 Hz), 5.34 (br d, 1H, J = 8.0 Hz), 5.03-4.92 (m, 2H) , 4.40 (m, 1H), 3.81 (d, d, 1H, J = 9.5, 2.9Hz), 3.74 (s, 3H), 3.61 (d, d, 1H, J = 9.5, 3.5Hz), 3.42 (m , 2H), 2.06 (quart., 2H, J = 7.3 Hz), 1.61 (quint., 2H, J = 7.3 Hz), 1.44 (s, 9 H).
Step 37B : Production of N-tert-butoxycarbonyl-O- (4-pentenyl) -L-serine

1.95 g (46 mmol) lithium hydroxide in 100 mL water in a solution of 6.65 g (23 mmol) N-tert-butoxycarbonyl-O- (4-pentenyl) -L-serine, methyl ester in 500 mL tetrahydrofuran at room temperature A monohydrate solution was added. The resulting mixture was stirred at room temperature for 40 hours. Next, 46 mL of 1.00N hydrochloric acid was added to the reaction mixture. The mixture was diluted with ethyl acetate, washed with water (3X) and brine, dried over anhydrous sodium sulfate, filtered then concentrated under reduced pressure to give 6.30 g of N-tert-butoxycarbonyl-O- (4- Pentenyl) -L-serine was obtained as a colorless oil. NMR (300 MHz, CDCl ₃ ): δ 5.77 (d d t, 1H, J = 17.2, 10.2, 6.6 Hz), 5.37 (br d, 1H, J = 8.0 Hz), 5.03-4.92 (m, 2H) , 4.42 (m, 1H), 3.87 (d d, 1H, J = 9.5, 2.6Hz), 3.63 (d d, 1H, J = 9.5, 4.0Hz), 3.45 (t, 2H, J = 6.6Hz ), 2.07 (quart., 2H, J = 7.3 Hz), 1.64 (quint., 2H, J = 7.3 Hz), 1.44 (s, 9 H).
Step 37C : (1S, 4R, 6S, 14S, 18R) -7-cis-14-tert-butoxycarbonylamino-18- (6-methoxyisoquinolin-1-yloxy) -2,15-dioxo-12-oxa- Preparation of 3,16-diazatricyclo [14.3.0.0 ^4,6 ] -nonadeca-7-ene-4-carboxylic acid (compound 84)

This compound was prepared from N-tert-butoxycarbonyl-3- (4-pentenylthio) -L-valine using the method described in Example 26. ESI-mass spectrum: m / e: 625 (M + H) ⁺ , 623 (MH) ⁻ ; High-resolution mass spectrum: Theoretical value for C ₃₂ H ₄₁ N ₄ O ₉ : 625.2874, Found: 625.22871; NMR (500 MHz , CD ₃ OD) δ 8.12 (d, 1H, J = 9.0Hz), 7.89 (d, 1H, J = 5.8Hz), 7.24 (d, 1H, J = 5.8Hz), 7.17 (s, 1H), 7.12 (d, 1H, J = 9.0Hz), 5.83 (s, 1H), 5.63 (quart., 1H, J = 8.8Hz), 5.45 (t, 1H, J = 8.8Hz), 4.74 (m, 1H), 4.60 (br s, 1H), 4.36 (d, 1H, J = 11.0Hz), 4.16 (d, 1H, J = 8.0Hz), 3.91 (s, 3H), 3.74 (m, 2H), 3.62 (m, 1H), 3.49 (m, 1H), 2.66 (m, 1H), 2.57 (m, 1H), 2.47-2.33 (m, 2H), 2.19 (m, 1H), 1.78 (m, 2H), 1.69 (m , 1H), 1.48 (m, 1H), 1.30 (s, 9H).
Step 37D : (1S, 4R, 6S, 14S, 18R)-[7-cis-4-cyclopropanesulfonyl-aminocarbonyl-18- (6-methoxy-isoquinolin-1-yloxy) -2,15-dioxo-12 -Oxa-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-en-14-yl] -carbamic acid, tert-butyl ester (compound 85)

This compound was prepared from compound 84 using the method described in Example 26I. ESI-mass spectrum: m / e: 728 (M + H) ⁺ , 726 (MH) ⁻ ; Theoretical value for high-resolution mass spectrum: C ₃₅ H ₄₆ N ₅ O ₁₀ S: 728.2966, found: 728.2972; NMR ( 500MHz, CD ₃ OD) δ 8.12 (m, 1H), 7.89 (d, 1H, J = 5.8Hz), 7.25 (d, 1H, J = 5.8Hz), 7.18 (s, 1H), 7.12 (d, 1H , J = 8.0Hz), 5.89 (s, 1H), 5.66 (quart., 1H, J = 8.8Hz), 5.19 (t, 1H, J = 8.8Hz), 4.71 (m, 1H), 4.52 (br s , 1H), 4.36 (d, 1H, J = 10.0Hz), 4.12 (m, 1H), 3.92 (s, 3H), 3.65 (m, 2H), 3.47 (m, 2H), 2.92 (m, 1H) 2.64 (m, 1H), 2.56 (m, 1H), 2.45 (m, 2H), 2.19 (m, 1H), 1.77 (m, 1H), 1.70-1.60 (m, 2H), 1.54 (m, 1H) ), 1.28 (s, 9H), 1.15-0.98 (m, 4H).
Example 38 : Preparation of compound 58:

工程38a：1-{[1-(2-tert-ブトキシカルボニルアミノ-ノナ-8-エノイル)-4-(tert-ブチル-ジメチル-シラニルオキシ)-ピロリジン-2-カルボニル]-アミノ}-2-ビニルシクロプロパンカルボン酸エチルエステルの製造

Step 38a : 1-{[1- (2-tert-butoxycarbonylamino-nona-8-enoyl) -4- (tert-butyl-dimethyl-silanyloxy) -pyrrolidine-2-carbonyl] -amino} -2-vinyl Production of cyclopropanecarboxylic acid ethyl ester

1-{[1- (2 (S) -tert-butoxycarbonylamino-nona-8-enoyl) -4 (R) -hydroxy-pyrrolidine-2 (S) carbonyl]-(1R) in 10 mL DMF -Amino} -2 (S) -vinyl-cyclopropanecarboxylic acid ethyl ester (1.5 g, 2.87 mmol) and imidazole (0.25 g, 3.67 mmol) and tert-butyl-dimethylsilyl chloride (516 mg, 3.44 mmol) Was added. The mixture was stirred at room temperature for 2 days. The reaction mixture was then concentrated under reduced pressure and the residue was dissolved in ethyl acetate. This solution was washed with water, dried over magnesium sulfate and then concentrated under reduced pressure to give a crude solid. Purify by flash chromatography (eluting with 20% ethyl acetate in hexanes) to 1.43 g (78%) of 1-{[1- (2-tert-butoxycarbonylamino-non-8-enoyl) -4- (tert -Butyl-dimethyl-silanyloxy) -pyrrolidine-2-carbonyl] -amino} -2-vinylcyclopropanecarboxylic acid ethyl ester was obtained as a white solid.
¹ H NMR (300 MHz, CD ₃ OD) δ 0.10 (s, 6H), 0.89 (s, 9H), 1.22 (m, 3H), 1.31-1.48 (m, 16H), 1.50-1.75 (m, 3H), 2.06 (m, 3H), 2.11-2.33 (m, 2H), 3.70 (m, 2H), 4.03-4.19 (m, 2H), 4.21 (m, 1H), 4.45 (t, J = 7.87Hz, 1H) , 4.59 (m, 1H), 4.91 (d, J = 9.15Hz, 1H), 4.98 (d, J = 17.20Hz, 1H), 5.08 (dd, J = 10.25, 1.83Hz, 1H), 5.27 (dd, J = 17.38, 1.65 Hz, 1H), 5.65-5.87 (m, 2 H). LC-MS (Method A, retention time: 4.00 min), MS m / z 636 (M ⁺ +1).
Step 38b : 14-tert-butoxycarbonylamino-18- (tert-butyl-dimethyl-silanyloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-ene- Production of 4-carboxylic acid, ethyl ester

1-{[1- (2-tert-Butoxycarbonylamino-non-8-enoyl) -4- (tert-butyl-dimethyl-silanyloxy) -pyrrolidine-2-carbonyl] -amino}-in 640 mL of methylene chloride To a solution of 2-vinyl-cyclopropanecarboxylic acid ethyl ester (1.63 g, 2.56 mmoL), 215 mg (0.26 mmoL) of tricyclohexylphosphine [1,3-bis (2,4,6-tri [benzylidene] ruthenium (IV The mixture was heated to reflux for 15 minutes and the residue was concentrated under reduced pressure and then purified by flash chromatography eluting with 30% ethyl acetate / hexanes. A second chromatography eluting with 50% ether in hexane gave 1.5 g (96%) of 14-tert-butoxycarbonylamino-18- (tert-butyl-dimethyl-silanyloxy) -2,15-dioxo-3, 16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-en-4- The carboxylic acid ethyl ester was obtained as a white solid: ¹ H NMR (500 MHz, CD ₃ Cl) δ 0.06 (s, 3H), 0.07 (s, 3H), 0.86 (s, 9H), 1.18-1.24 (m, 6H ), 1.34-1.64 (m, 14H), 1.86-1.96 (m, 3H), 2.02-2.09 (m, 1H), 2.11-2.17 (m, 1H), 2.19-2.28 (m, 1H), 2.57-2.63 (m, 1H), 3.50-3.54 (m, 1H), 3.71 (dd, J = 10.22, 6.26Hz, 1H), 4.06-4.17 (m, 2H), 4.52-4.58 (m, 2H), 4.75 (d , J = 8.55Hz, 1H), 5.21 (t, J = 9.92Hz, 1H), 5.35 (d, J = 7.63Hz, 1H), 5.45-5.50 (m, 1H), 6.94 (s, 1H) .LC -MS (Method A, retention time: 3.88 min), MS m / z 608 (M ⁺ +1).
Step 38c : 14-tert-butoxycarbonylamino-18- (tert-butyl-dimethyl-silanyloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-ene- 4-carboxylic acid production

14-tert-butoxycarbonylamino-18- (tert-butyl-dimethyl-silanyloxy) -2,15-dioxo-3, in a mixed solvent system of THF (4 mL), methanol (1 mL), and water (2 mL), Powdered lithium hydroxide monohydrate (1.0 g, 50 mmoL) in a solution of 16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-ene-4-carboxylic acid ethyl ester (1.5 g, 2.47 mmoL) Was added. The pale yellow slurry was stirred at room temperature under N ₂ for 4 hours. The mixture was then concentrated under reduced pressure and the residue was partitioned between ether and water. The ether phase was discarded and the aqueous phase was treated with 1N HCl until pH 4 was reached. The acidic solution was extracted with EtOAc (3x). The combined EtOAc extracts were dried (MgSO ₄ ) and then concentrated under reduced pressure. 1.2 g (84%) 14-tert-butoxycarbonylamino-18- (tert-butyl-dimethyl-silanyloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca- 7-ene-4-carboxylic acid was obtained as an off-white solid. ¹ H NMR (300 MHz, CD ₃ OD) 0.12 (s, 6H), 0.89 (s, 9H), 1.23-1.64 (m, 17H), 1.70-1.87 (m, 1H), 1.90-2.49 (m, 6H) , 3.70-3.80 (m, 1H), 3.83-3.90 (m, 1H), 4.28-4.36 (m, 1H), 4.47-4.55 (m, 1H), 4.65 (s, 1H), 5.30-5.39 (m, 1H), 5.53-5.62 (m, 1H). LC-MS (Method A, retention time: 3.69 min), MS m / z 580 (M ⁺ +1).
Step 38d : [18- (tert-butyl-dimethyl-silanyloxy) -4-cyclopropanesulfonylaminocarbonyl-2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-ene Of -14-yl] -carbamic acid tert-butyl ester

14-tert-Butoxycarbonylamino-18- (tert-butyl-dimethyl-silanyloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-ene-4-carboxylic acid The acid (500 mg, 0.86 mmoL) was dissolved in 25 mL THF and then treated with CDI (180 mg, 1.12 mmoL). (Care was taken to avoid moisture using an oven-dried glass container and maintaining a dry N ₂ atmosphere) The reaction mixture was refluxed for 2 hours, then cooled to room temperature and then sequentially treated with cyclopropylsulfonamide (135 mg, 1.12 mmoL) and DBU (170 mg, 1.12 mmoL). The reaction mixture was stirred at room temperature for 4 hours and then the THF was removed by rotary evaporation. The residue was partitioned between ethyl acetate and pH 4 buffer. The organic layer was dried (MgSO ₄ ) and then concentrated under reduced pressure to give the crude product. It was then purified by flash chromatography (eluting with 33% ethyl acetate in hexane) to give 300 mg (51%) of [18- (tert-butyl-dimethyl-silanyloxy) -4-cyclopropanesulfonylaminocarbonyl-2 , 15-Dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-en-14-yl] -carbamic acid tert-butyl ester was obtained as a white solid. ¹ H NMR (300 MHz, CD ₃ OD) δ 1H 0.07 (s, 3H), 0.08 (s, 3H), 0.85 (s, 9H), 0.87-1.49 (m, 21H), 1.73-1.95 (m, 3H) 2.08-2.16 (m, 1H), 2.25-2.36 (m, 2H), 2.42-2.56 (m, 1H), 2.85-2.93 (m, 1H), 3.65-3.74 (dd, J = 10.61, 3.66Hz, 1H), 3.89 (d, J = 10.25Hz, 1H), 4.34 (m, J = 9.70, 9.70Hz, 1H), 4.43 (t, J = 7.87Hz, 1H), 4.57 (s, 1H), 4.94- 5.01 (m, 1H), 5.10 (d, J = 8.78Hz, 1H), 5.66-5.75 (m, 1H), 6.55 (s, 1H), 10.13 (s, 1H). LC-MS (Method A, retention Time: 3.81 min), MS m / z 683 (M ⁺ +1).
Step 38e : (4-Cyclopropanesulfonylaminocarbonyl-18-hydroxy-2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-en-14-yl) -carbamic acid tert-butyl ester

[18- (tert-Butyl-dimethylsilanyloxy) -4-cyclopropanesulfonylaminocarbonyl-2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca- in 25 mL THF To a mixture of 7-en-14-yl] -carbamic acid tert-butyl ester (330 mg, 0.48 mmoL) was added tetrabutylammonium fluoride (150 mg, 0.54 mmoL). The reaction mixture was stirred at room temperature for 18 hours and then the THF was removed by rotary evaporation. The residue was partitioned between ethyl acetate and water. The organic layer was dried (MgSO ₄ ) and then concentrated under reduced pressure to give the crude product. Next, triturated with hexane, 200 mg (73%) of (4-cyclopropanesulfonylaminocarbonyl-18-hydroxy-2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca- 7-en-14-yl) -carbamic acid tert-butyl ester was obtained as a white solid. ¹ H NMR (500 MHz, CD ₃ Cl) δ 1.87-1.64 (m, 21H), 1.70-1.98 (m, 3H), 2.15-2.56 (m, 5H), 2.85-2.94 (m, 1H), 3.71 (d , J = 13.91Hz, 1H), 4.10-4.26 (m, 2H), 4.51 (t, J = 7.87Hz, 1H), 4.62 (s, 1H), 4.98 (m, 1H), 5.06 (d, J = 8.78Hz, 1H), 5.64-5.71 (m, 1H), 6.72 (s, 1H), 10.24 (s, 1H). LC-MS (Method A, retention time: 2.85min), MS m / z 569 (M ⁺ +1).
Step 38f : Compound 58: [(Z)-(1S, 4R, 14S, 18R) -4-cyclopropanesulfonylaminocarbonyl-2,15-dioxo-18- (3-phenyl-isoquinolin-1-yloxy) -3 Of 1,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-en-14-yl] -carbamic acid tert-butyl ester

(4-Cyclopropanesulfonylaminocarbonyl-18-hydroxy-2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-en-14-yl) in 1 mL DMF To a mixture of carbamic acid tert-butyl ester (20 mg, 0.035 mmol), potassium t-butoxide (22 mg, 0.196 mmol) was added. The mixture was stirred at room temperature for 5 minutes and then 1-chloro-3-phenylisoquinoline (15 mg, 0.062 mmol) was added. The reaction mixture was stirred at room temperature for 15 hours and then concentrated under reduced pressure. The crude product was triturated with ether. The residue was dissolved in MeOH and then purified by preparative HPLC. (YMC XTERRA, S5, 19x100 mm, gradient: 60% B-100% B, 15 min, retention 2 min, flow rate 25 mL / min) to 10 mg (39%) Compound 58, [(Z)-(1S, 4R, 14S, 18R) -4-cyclopropanesulfonylaminocarbonyl-2,15-dioxo-18- (3-phenyl-isoquinolin-1-yloxy) -3,16- Diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-en-14-yl] -carbamic acid tert-butyl ester was obtained as a white solid. ¹ H NMR (500 MHz, CD ₃ Cl) δ 0.90-1.64 (m, 21H), 1.76-1.97 (m, 3H), 2.26-2.31 (m, 1H), 2.52-2.63 (br s, 1H), 2.69- 2.81 (m, 2H), 2.88-2.93 (m, 1H), 4.11 (d, J = 11.60Hz, 1H), 4.33 (m, 1H), 4.61 (d, J = 7.94Hz, 2H), 4.99 (t , J = 9.31Hz, 1H), 5.05 (d, J = 7.93Hz, 1H), 5.69-5.74 (m, 1H), 6.08 (s, 1H), 6.60 (s, 1H), 7.38-7.47 (m, 2H), 7.50 (t, J = 7.63Hz, 2H), 7.63 (t, J = 7.32Hz, 1H), 7.71 (s, 1H), 7.77 (d, J = 8.24Hz, 1H), 8.10 (d, . J = 7.32Hz, 2H), 8.18 (d, J = 7.93Hz, 1H), 10.27 (s, 1H) LC-MS ( method A, retention time: 3.72min), MS m / z 772 (M + + 1).
Example 39 : Preparation of compounds 144 and 145

工程39a：(S)-2-(tert-ブトキシカルボニル)-3-(2-ニトロ-N-(ペンタ-4-エニル)フェニルスルホンアミド)プロピオン酸vの製造

Step 39a : Preparation of (S) -2- (tert-butoxycarbonyl) -3- (2-nitro-N- (pent-4-enyl) phenylsulfonamido) propionic acid v

Preparation of (S) -methyl 3-amino-2- (tert-butoxycarbonyl) propionate ii:

To a mixture of i (Boc-DAP-OH) (3.0 g 14.7 mmol) in 50 mL of methylene chloride was added 5 mL of methanol. To this solution (trimethylsilyl) diazomethane (2M in ether, 7.9 mL, 15.8 mmol) was added slowly. The mixture was stirred at room temperature for 2 hours until all solids were dissolved and the solution was pale yellow. It was then concentrated to give 3.2 g (99%) of (S) -methyl 3-amino-2- (tert-butoxycarbonyl) propionate ii as a colorless oil. ¹ H NMR (CD ₃ OD, 300 MHz) δ 1.46 (s, 9H), 2.82-3.00 (m, 2H), 3.71 (s, 3H), 4.14 (brs, 1H).
Preparation of (S) -methyl 2- (tert-butoxycarbonyl) -3- (2-nitrophenylsulfonamido) propionate iii:

To a mixture of (S) -methyl 3-amino-2- (tert-butoxycarbonyl) propionate ii (1.6 g, 7.3 mmol) in DCM (50 mL) was added DIPEA (1.64 mL, 9.4 mmol) and 2-nitrobenzenesulfonyl chloride. (1.62 g, 7.3 mmoL) was added. The mixture was stirred at room temperature for 2 hours. It was then concentrated and dissolved in ethyl acetate, which was then washed with sat. Sodium bicarbonate, brine and then dried over magnesium sulfate. It was then filtered and concentrated to give 2.9 g (98%) of (S) -methyl 2- (tert-butoxycarbonyl) -3- (2-nitrophenylsulfonamido) propionate iii as a yellow foam. Obtained. ¹ H NMR (CD ₃ OD, 300 MHz) δ 1.41 (s, 9H), 3.36-3.51 (m, 2H), 3.71 (s, 3H), 4.22 (m, 1H), 7.80-7.90 (m, 3H), 8.07-8.10 (m, 1H).
Preparation of (S) -methyl 2- (tert-butoxycarbonyl) -3- (2-nitro-N- (pent-4-enyl) phenylsulfonamido) propionate iv:

To a mixture of (S) -methyl 2- (tert-butoxycarbonyl) -3- (2-nitrophenylsulfonamido) -propionate iii (150 mg, 0.37 mmol) in 3 mL DMF was added potassium carbonate (102 mg, 0.74 mmol). ) Was added. The mixture was stirred at room temperature for 20 minutes and then 5-bromo-1-pentene (65 μL, 0.55 mmoL) was added. The reaction mixture was stirred at room temperature for 2 days. It was then filtered, concentrated and then purified by silica gel chromatography (eluting with 25% ethyl acetate in hexane) to yield 75 mg (43%) of (S) -methyl 2- (tert-butoxycarbonyl) -3. -(2-Nitro-N- (pent-4-enyl) phenylsulfonamido) propionate iv was obtained as a yellow solid. ¹ H NMR (CD ₃ OD, 300 MHz) δ 1.42 (s, 9H), 1.54-1.64 (m, 2H), 1.97 (q, J = 7.20Hz, 2H), 3.37 (m, 2H), 3.57-3.80 ( m, 2H), 3.72 (s, 3H), 4.42 (dd, J = 8.60, 5.31Hz, 1H), 4.91-5.01 (m, 2H), 5.69-5.79 (m, 1H), 7.75-7.85 (m, 3H), 8.04 (m, 1H). LC-MS (Method D, retention time: 2.68 min), MS m / z 372 (M ⁺ + 1-Boc).
Preparation of (S) -2- (tert-butoxycarbonyl) -3- (2-nitro-N- (pent-4-enyl) phenylsulfonamido) propionic acid v :

(S) -methyl 2- (tert-butoxycarbonyl) -3- (2-nitro-N- (pent-4-enyl) phenylsulfonamido) -propionate iv (500 mg, 1.06 mmol) mixed solvent system: THF ( 4 mL), methanol (1 mL), and water (2 mL). Powdered lithium hydroxide (250 mg, 10.4 mmol) was added. The pale yellow slurry was stirred at room temperature for 15 hours and then concentrated under reduced pressure. The residue was partitioned between ether and water. The ether phase was discarded and the aqueous phase was treated with 1N HCl until the pH was 4. The acidic solution was extracted with ethyl acetate for 4 hours. The combined ethyl acetate extracts were dried (MgSO ₄ ) and then concentrated under reduced pressure to yield 430 mg (89%) of (S) -2- (tert-butoxycarbonyl) -3- (2-nitro-N- (penta). -4-enyl) phenylsulfonamido) propionic acid v was obtained as a yellow oil. ¹ H NMR (CD ₃ OD, 300 MHz) δ 1.38 (s, 9H), 1.51-1.60 (m, 2H), 1.89-1.98 (m, 2H), 3.28-3.32 (m, 2H), 3.59-3.64 (dd , J = 14.95, 9.46Hz, 1H), 3.71-3.74 (m, 1H), 4.33 (dd, J = 9.61, 4.43Hz, 1H), 4.87-4.94 (m, 2H), 5.63-5.72 (m, 1H) ), 7.71-7.77 (m, 3H), 8.01 (dd, J = 7.48, 1.37Hz, 1H). LC-MS (Method D, retention time: 2.04 min), MS m / z 358 (M ⁺ + 1- Boc).
Step 39b : (1R, 2S) -ethyl 1-((3R, 5S) -1-((S) -2- (tert-butoxycarbonyl) -3- (2-nitro-N- (pent-4-enyl) Preparation of) phenylsulfonamido) propanoyl) -3- (isoquinolin-1-yloxy) pyrrolidine-5-carboxamide) -2-vinylcyclopropanecarboxylate

(S) -2- (tert-butoxycarbonyl) -3- (2-nitro-N- (pent-4-enyl) phenylsulfonamido) propionic acid (1.10 g, 2.40 mmol) dissolved in 20 mL DCM was added. Sequentially 1-{[4- (isoquinolin-1-yloxy) -pyrrolidine-2-carbonyl] -amino} -2-vinyl-cyclopropanecarboxylic acid ethyl ester, bishydrochloric acid (1.27 g, 2.72 mmol), N-methylmorpholine (0.92 mL, 8.34 mmol), and HATU (PE biosystems) (1.28 g, 3.36 mmol). The reaction mixture was stirred at room temperature under N ₂ for 15 hours and then concentrated under reduced pressure. The residue was partitioned between ethyl acetate and pH 4 buffer (biphthalate). The organic layer was washed with sat. Aq. NaHCO ₃ , dried (MgSO ₄ ) and then concentrated under reduced pressure to give 2.0 g of crude product. By flash chromatography (30% hexane / ethyl acetate) 1.4 g (70%) of (1R, 2S) -ethyl 1-((3R, 5S) -1-((S) -2- (tert-butoxycarbonyl)- 3- (2-Nitro-N- (pent-4-enyl) phenylsulfonamido) propanoyl) -3- (isoquinolin-1-yloxy) pyrrolidine-5-carboxamide) -2-vinylcyclopropanecarboxylate as a purple solid Obtained. ¹ H NMR (CD ₃ OD, 500 MHz) δ 1.23-1.31 (m, 9H), 1.46-1.52 (m, 4H), 1.59 (br s, 1H), 1.66 (br s, 1H), 1.74 (dd, J = 7.93, 5.19Hz, 1H), 2.00 (br s, 2H), 2.29 (q, J = 8.95Hz, 1H), 2.47 (m, 1H), 2.72 (m, 1H), 3.38-3.51 (m, 2H) ), 3.68 (m, 2H), 4.09-4.26 (m, 3H), 4.37 (d, J = 11.29Hz, 1H), 4.64 (m, 1H), 4.72 (m, 1H), 4.95 (d, J = 10.99Hz, 1H), 4.99 (dd, J = 18.92, 1.53Hz, 1H), 5.12 (d, J = 10.68Hz, 1H), 5.32 (d, J = 17.40Hz, 1H), 5.73-5.86 (m, 2H), 5.92 (s, 1H), 7.36 (m, 1H), 7.58 (m, 1H), 7.73 (m, 1H), 7.80-7.88 (m, 4H), 7.99 (m, 1H), 8.11 (m , 1H), 8.24 (d, J = 8.24 Hz, 1H). LC-MS (Method D, retention time: 3.15 min), MS m / z 835 (M ⁺ +1).
Step 39c :

(1R, 2S) -Ethyl 1-((3R, 5S) -1-((S) -2- (tert-butoxycarbonyl) -3- (2-nitro-N- (penta- 4-enyl) phenylsulfonamido) propanoyl) -3- (isoquinolin-1-yloxy) pyrrolidine-5-carboxamide) -2-vinylcyclopropanecarboxylate (1.40 g, 1.67 mmol) and tricyclohexylphosphine [1,3- A solution of bis (2,4,6-trimethylphenyl) -4,5-dihydroimidazol-2-ylidene] [benzylidene] ruthenium (IV) dichloride (Aldrich), (204 mg, 0.24 mmol) was refluxed under N ₂ . . Immediately after reaching the reflux temperature, the reaction mixture was homogeneous and pale orange. After refluxing for 2 hours, the dark orange reaction mixture was cooled to room temperature and then concentrated under reduced pressure to give 1.7 g of an orange oil. Flash chromatography (30% hexane / ethyl acetate) gave 1.1 g (82%) of the above ester as a purple solid. ¹ H NMR (CD ₃ OD, 300 MHz) δ 1.11-1.31 (m, 12H), 1.51-1.80 (m, 4H), 1.97-2.15 (m, 2H), 2.34-2.50 (m, 2H), 2.62-2.69 (m, 1H), 3.17-3.33 (m, 2H), 3.44-3.60 (m, 2H), 4.01-4.05 (m, 1H), 4.12 (q, J = 7.32Hz, 2H), 4.37 (d, J = 13.17Hz, 1H), 4.58 (t, J = 8.42Hz, 1H), 4.71 (m, 1H), 5.61 (m, 2H), 5.88 (s, 1H), 6.91 (d, J = 9.15Hz, 1H ), 7.30 (d, J = 5.49Hz, 1H), 7.52 (t, J = 7.50Hz, 1H), 7.66-7.73 (m, 1H), 7.76-7.80 (m, 4H), 7.95 (d, J = 5.86Hz, 1H), 8.11 (d, J = 7.32Hz, 1H), 8.17 (d, J = 8.05Hz, 1H), 8.84 (s, 1H). LC-MS (Method D, retention time: 2.91min) MS m / z 807 (M ⁺ +1).
Step 39d : Production of Compound 144

The ethyl ester prepared in step 39c (1.1 g, 1.3 mmol) was dissolved in a mixed solvent system: THF (16 mL), methanol (4 mL), and water (8 mL). Powdered lithium hydroxide hydrate (540 mg, 13.2 mmol) was added. The pale yellow slurry was stirred at room temperature for 15 hours and then concentrated under reduced pressure. The residue was partitioned between ether and water. The ether phase was discarded and the aqueous phase was treated with 1N HCl until the pH was 4. The acidic solution was extracted 4 times with ethyl acetate. The combined ethyl acetate extracts were dried (MgSO ₄ ) and then concentrated under reduced pressure to give 878 mg (85%) of compound 144 as a purple solid. ¹ H NMR (CD ₃ OD, 300 MHz) δ 1.11, 1.31 (s, 9H), 1.56-1.68 (m, 4H), 2.04-2.16 (m, 2H), 2.42-2.66 (m, 2H), 2.68-2.75 (m, 1H), 3.22-3.30 (m, 2H), 3.48-3.63 (m, 2H), 4.02-4.14 (m, 1H), 4.46 (d, J = 12.44Hz, 1H), 4.62 (t, J = 8.42Hz, 1H), 4.70-4.73 (m, 1H), 5.64 (m, 2H), 5.90 (s, 1H), 7.36 (d, J = 5.86Hz, 1H), 7.57 (t, J = 7.68Hz , 1H), 7.71-7.87 (m, 5H), 7.98 (d, J = 5.86Hz, 1H), 8.15 (m, 1H), 8.22 (d, J = 8.05Hz, 1H). LC-MS (Method D , Retention time: 2.78 min), MS m / z 779 (M ⁺ +1).
Step 39e : Production of Compound 145

Compound 144 (0.878 g, 1.1 mmol) was dissolved in 15 mL of THF and treated with CDI (255 mg, 1.58 mmol). (Care was taken to avoid moisture by using an oven-dried glass container and to maintain a dry N ₂ atmosphere.) After refluxing the reaction mixture for 1 hour, it was cooled to room temperature and then sequentially cyclopropylsulfonamide (194 mg, 1.6 mmol) and DBU (245 mg, 1.6 mmol) were added. After stirring at room temperature for 24 hours, THF was removed by rotary evaporation. The residue was partitioned between ethyl acetate and pH 4 buffer. The organic layer was dried (MgSO ₄ ) and then concentrated under reduced pressure to give the crude product. Flash chromatography (3% MeOH / methylene chloride) gave 0.68 g (70%) of compound 145 as a white solid. ¹ H NMR (CD ₃ OD, 300 MHz) δ 1.07-1.37 (m, 13H), 1.11 (s, 9H), 1.60-1.78 (m, 4H), 1.97-2.02 (m, 1H), 2.28-2.35 (m , 1H), 2.44-2.51 (m, 1H), 2.75-2.68 (m, 1H), 2.75-2.80 (m, 1H), 2.94-3.00 (m, 1H), 3.20-3.25 (m, 1H), 3.28 -3.47 (m, 2H), 3.57-3.62 (m, 1H), 4.05-4.08 (m, 1H), 4.51 (d, J = 12.21Hz, 1H), 4.65-4.68 (m, 1H), 4.84 (m , 1H), 5.15-5.23 (m, 1H), 5.70-5.75 (m, 1H), 5.94 (s, 1H), 7.35 (d, J = 6.10Hz, 1H), 7.57 (t, J = 7.32Hz, 1H), 7.74 (t, J = 7.78Hz, 1H), 7.78-7.89 (m, 4H), 8.00 (d, J = 5.80Hz, 1H), 8.20-8.25 (m, 2H). LC-MS ( Method D, retention time: 2.89 min), MS m / z 882 (M ⁺ +1).
Example 40 : Preparation of compound 151

To a mixture of compound 145 (680 mg, 0.77 mmol) in acetonitrile (20 mL) was added potassium carbonate (319 mg, 2.31 mmol) and benzenethiol (186 mg, 1.69 mmol). The mixture was stirred at room temperature overnight and then concentrated under reduced pressure. The residue was diluted with water and the aqueous solution was then adjusted to pH 5 with 1N HCl. A precipitated white solid was formed in the solution. This heterogeneous mixture was extracted with ethyl acetate (3x). The combined extract was dried (MgSO ₄ ) and then concentrated to give a pale yellow solid. The crude solid was then triturated several times with hexanes to give 510 mg (95%) of compound 151 as an off-white solid. ¹ H NMR (CD ₃ OD, 300 MHz) δ 1.06-1.88 (m, 13H), 1.23, 1.31 (s, 9H), 1.65 (dd, J = 9.70, 5.67 Hz, 1H), 1.80 (dd, J = 8.23 , 5.67Hz, 2H), 2.03 (brs, 1H), 2.18 (br s, 1H), 2.37-2.52 (m, 3H), 2.85-2.99 (m, 2H), 3.09-3.15 (m, 1H), 3.27 (m, 1H), 3.44 (m, 2H), 4.15-4.20 (m, 1H), 4.46 (m, 1H), 4.74-4.85 (m, 2H), 5.32 (t, J = 10.25Hz, 1H), 5.71 (m, 1H), 5.98 (br s, 1H), 7.37 (d, J = 5.86Hz, 1H), 7.58 (t, J = 8.23Hz, 1H), 7.74 (t, J = 7.68Hz, 1H) , 7.85 (d, J = 8.05Hz, 1H), 8.00 (d, J = 5.86Hz, 1H), 8.22 (d, J = 8.05Hz, 1H). LC-MS (Method D, retention time: 2.01min) MS m / z 697 (M ⁺ +1).
Example 41 : Preparation of compound 146

To a mixture of compound 151 (20 mg, 0.029 mmol) in 2 mL DCM was added triethylamine (14 mg, 0.138 mmol) and acetic anhydride (8 mg, 0.078 mmol). The reaction mixture was stirred at room temperature for 2 hours and then concentrated under reduced pressure. The residue was dissolved in methanol and then purified by preparative HPLC (YMC XTERRA, S5, 30x50mm, gradient: 45% B to 85% B, 15 min, retention 2 min, flow rate 25 mL / min) to give 10 mg (47%) of compound 146 was obtained as a white solid. ¹ H NMR (CD ₃ OD, 300 MHz, 60 ° C.) δ 1.08-1.22 (m, 13H), 1.55-1.74 (m, 4H), 2.09-2.21 (m, 2H), 2.13 (s, 3H), 2.33- 2.48 (m, 2H), 2.66-2.75 (m, 1H), 2.90-2.99 (m, 1H), 3.12-3.25 (m, 2H), 3.45-3.60 (m, 2H), 3.75-3.84 (m, 1H ), 4.98-4.05 (m, 1H), 4.70 (t, J = 7.68Hz, 1H), 4.89-4.97 (m, 1H), 5.26 (t, J = 10.43Hz, 1H), 5.2-5.75 (m, 1H), 5.92 (s, 1H), 7.29 (d, J = 5.86Hz, 1H), 7.50 (t, J = 7.50Hz, 1H), 7.64-7.69 (m, 1H), 7.77 (m, 1H), 7.95 (d, J = 5.86 Hz, 1H), 8.18 (d, J = 8.78 Hz, 1H). LC-MS (Method D, retention time: 2.64 min), MS m / z 739 (M ⁺ +1).
Example 42 : Preparation of compound 147

To a mixture of compound 151 (100 mg, 0.144 mmol) in 5 mL of DMF was added potassium carbonate (40 mg, 0.290 mmol) and methyl iodide (13 μL, 0.21 mmol). The reaction mixture was stirred at room temperature for 2 hours. It was then concentrated under reduced pressure. The residue was dissolved in methanol and purified by preparative HPLC (YMC XTERRA, S5, 30x50 mm, gradient: 45% B to 70% B, 15 min, retention 2 min, flow rate 25 mL / min) to give 75 mg (73%) of compound 147 Was obtained as a white solid. ¹ H NMR (CD ₃ OD, 300 MHz, 60 ° C.) δ 1.02-1.28 (m, 13H), 1.64 (dd, J = 9.70, 5.67 Hz, 1H), 1.72-1.81 (m, 2H), 1.91-2.10 ( m, 1H), 2.13-2.30 (m, 1H), 2.36-2.50 (m, 2H), 2.54-2.68 (m, 1H), 2.76-2.99 (m, 2H), 3.05 (s, 3H), 3.11- 3.46 (m, 4H), 3.79-3.96 (m, 1H), 4.17 (m, 1H), 4.76 (m, 2H), 5.31 (t, J = 10.06Hz, 1H), 5.71 (m, 1H), 5.99 (s, 1H), 7.33 (d, J = 5.86Hz, 1H), 7.55 (t, J = 7.50Hz, 1H), 7.70 (t, J = 7.68Hz, 1H), 7.81 (m, 1H), 7.96 (d, J = 5.86 Hz, 1H), 8.17 (d, J = 8.42 Hz, 1H). LC-MS (Method D, retention time: 1.98 min), MS m / z 711 (M ⁺ +1).
Example 43 : Preparation of compound 148

To a mixture of compound 151 (20 mg, 0.029 mmol) in 2 mL DCM was added diisopropylamine (15 μL, 0.086 mmol) and methyl chloroformate (8 mg, 0.085 mmol). The reaction mixture was stirred at room temperature for 2 hours and then concentrated under reduced pressure. The residue was dissolved in methanol and then purified by preparative HPLC (YMC XTERRA, S5, 30x50 mm, gradient: 50% B to 90% B, 15 min, retention 2 min, flow rate 25 mL / min) to give 15 mg (69%) of compound 148 was obtained as a white solid. ¹ H NMR (CD ₃ OD, 300 MHz) δ 1.09 (s, 9H), 1.07-1.45 (m, 13H), 1.56-1.61 (m, 1H), 1.70-1.76 (m, 3H), 2.95-2.10 (m , 1H), 2.13-2.26 (m, 1H), 2.33-2.55 (m, 2H), 2.71-2.78 (m, 1H), 2.93-3.02 (m, 1H), 3.33 (m, 2H), 3.48-3.57 (m, 2H), 3.75 (s, 3H), 4.04-4.12 (m, 1H), 4.64 (t, J = 8.42Hz, 1H), 4.90 (m, 2H), 5.19 (t, J = 10.25Hz, 1H), 5.69-5.77 (m, 1H), 5.93 (s, 1H), 7.34 (d, J = 5.86Hz, 1H), 7.55 (t, J = 6.95Hz, 1H), 7.72 (t, J = 7.32 Hz, 1H), 7.83 (d, J = 7.32Hz, 1H), 7.98 (d, J = 5.86Hz, 1H), 8.20 (d, J = 8.42Hz, 1H), 9.21 (s, 1H) .LC- MS (Method D, retention time: 2.75 min), MS m / z 755 (M ⁺ +1).
Example 44 : Preparation of compound 150

To a mixture of compound 151 (20 mg, 0.029 mmol) in 2 mL DCM was added diisopropylamine (15 μL, 0.086 mmol) and tert-butyl isocyanate (8 μL, 0.058 mmol). The reaction mixture was stirred at room temperature for 2 hours and then concentrated under reduced pressure. The residue was dissolved in methanol and then purified by preparative HPLC (YMC XTERRA, S5, 30x50mm, gradient: 55% B to 90% B, 15 min, retention 2 min, flow rate 25 mL / min) to give 15 mg (65%) of compound 150 was obtained as a white solid (15 mg, 65%). ¹ H NMR (CD ₃ OD, 300 MHz) δ 1.06-1.34 (m, 13H), 1.21 (s, 9H), 1.40 (s, 9H), 1.58-1.75 (m, 3H), 1.97-2.09 (m, 1H ), 2.11-2.23 (m, 1H), 2.37-2.50 (m, 2H), 2.69-2.82 (m, 1H), 2.94-3.04 (m, 1H), 3.22 (m, 1H), .36-3.46 ( m, 2H), 4.12 (d, J = 12.44Hz, 1H), 4.39 (d, J = 11.34Hz, 1H), 4.50-4.57 (m, 1H), 4.69 (t, J = 8.23Hz, 1H), 5.20-5.27 (m, 1H), 5.69-5.78 (m, 1H), 5.90 (s, 1H), 7.34 (d, J = 5.86Hz, 1H), 7.57 (t, J = 7.14Hz, 1H), 7.73 (m, 1H), 7.83 (m, 1H), 7.98 (d, J = 5.86Hz, 1H), 8.24 (d, J = 8.05Hz, 1H). LC-MS (Method D, retention time: 3.04min) MS m / z 796 (M ⁺ +1).
Example 45 : Preparation of compound 153

To a mixture of compound 151 (20 mg, 0.029 mmol) in 2 mL DCM was added diisopropylamine (15 μL, 0.086 mmol) and methanesulfonyl chloride (5 μL 0.065 mmol). The reaction mixture was stirred at room temperature for 2 hours and then concentrated under reduced pressure. The residue was dissolved in methanol and then purified by preparative HPLC (YMC XTERRA, S5, 30x50 mm, gradient: 50% B to 90% B, 15 min, retention 2 min, flow rate 25 mL / min) to give 17 mg (76%) of compound 153 was obtained as a white solid. ¹ H NMR (CD ₃ OD, 300 MHz) δ 1.02-1.49 (m, 13H), 1.59 (m, 1H), 1.72-1.86 (m, 3H), 1.96-2.09 (m, 1H), 2.37-2.50 (m , 2H), 2.63-2.80 (m, 2H), 2.91-3.08 (m, 5H), 2.97 (s, 3H), 3.22-3.45 (m, 3H), 4.19 (dd, J = 11.89, 3.48Hz, 1H ), 4.52 (d, J = 11.71Hz, 1H), 4.62 (dd, J = 9.88, 7.32Hz, 1H), 4.83-4.96 (m, 1H), 5.17 (t, J = 10.06Hz, 1H), 5.75 (m, 1H), 5.94 (s, 1H), 7.33 (d, J = 5.86Hz, 1H), 7.57 (d, J = 8.05Hz, 1H), 7.72 (t, J = 7.50Hz, 1H), 7.82 (m, 1H), 7.99 (d, J = 6.22Hz, 1H), 8.20 (d, J = 8.42Hz, 1H). LC-MS (Method D, retention time: 2.59min), MS m / z 775 ( M ⁺ +1).
Example 46 Compound 141: [(Z)-(1S, 4R, 13S, 17R) -4-cyclopropanesulfonylaminocarbonyl-17- (6-methoxy-isoquinolin-1-yloxy) -2,14-dioxo- Preparation of 10-oxa-3,15-diaza-tricyclo [13.3.0.0 ^4,6 ] octadeca-7-en-13-yl] -carbamic acid tert-butyl ester

工程46a：(S)-4-アリルオキシ-2-(tert-ブトキシカルボニルアミノ)酪酸の製造

Step 46a : Production of (S) -4-allyloxy-2- (tert-butoxycarbonylamino) butyric acid

To a mixture of sodium hydride (913 mg, 22.8 mmoL) in DMF at 0 ° C. was added Nt-Boc-L-homoserine (2 g, 9.13 mmoL). The reaction mixture was stirred at 0 ° C. for 15 minutes and then allyl bromide (1.38 g, 11.4 mmoL) was added. The mixture was warmed to room temperature and stirred for 2 hours. It was then concentrated under reduced pressure. The residue was diluted with water and then washed sequentially with hexane and ether. The organic layer was discarded and the aqueous layer was carefully adjusted to pH 3 with 1N HCl. This acidic aqueous solution was extracted with ethyl acetate. The organic layer was dried (MgSO ₄ ) and then concentrated under reduced pressure to give 2.2 g (93%) of (S) -4-allyloxy-2- (tert-butoxycarbonylamino) butyric acid as a colorless oil. ¹ H NMR (300 MHz, CD ₃ OD) δ 1.42 (s, 9H), 1.80-1.90 (m, 1H), 2.04-2.16 (m, 1H), 3.50-3.54 (m, 2H), 3.97 (d, J = 4.39Hz, 2H), 4.23 (dd, J = 8.78, 4.39Hz, 1H), 5.15 (d, J = 10.25Hz, 1H), 5.26 (dd, J = 17.38, 1.65Hz, 1H), 5.84-5.97 (m, 1H). This starting material was (1R, 2S) -ethyl 1-((3R, 5S) -1-((S) -4- (allyloxy) -2- (tert-butoxycarbonyl) butanoyl) as described in Example 2. Used in the synthesis of -3- (6-methoxyisoquinolin-1-yloxy) pyrrolidine-5-carboxamide) -2-vinylcyclopropane-carboxylate.
Step 46b : (Z)-(1S, 4R, 13S, 17R) -13-tert-butoxycarbonylamino-17- (6-methoxy-isoquinolin-1-yloxy) -2,14-dioxo-10-oxa-3 Of 1,15-diaza-tricyclo [13.3.0.0 ^4,6 ] octadeca-7-ene-4-carboxylic acid ethyl ester

(1R, 2S) -Ethyl 1-((3R, 5S) -1-((S) -4- (allyloxy) -2- (tert-butoxycarbonyl) butanoyl) -3- (6 -Methoxyisoquinolin-1-yloxy) pyrrolidine-5-carboxamide) -2-vinylcyclopropanecarboxylate (220 mg, 0.33 mmol) in a solution of tricyclohexylphosphine [1,3-bis (2,4,6-trimethyl-phenyl) ) -4,5-dihydroimidazol-2-ylidene] [benzylidene] -ruthenium (IV) dichloride (27 mg, 0.032 mmol) was obtained. The pale orange homogeneous solution was refluxed for 3 hours to give a dark orange solution. At this time, another portion of ruthenium catalyst (13 mg, 0.016 mmol) was added and then reflux continued for another 2 hours. The reaction mixture was cooled to room temperature and then concentrated under reduced pressure to give an orange oil. 174 mg (80%) (Z)-(1S, 4R, 13S, 17R) -13-tert-butoxycarbonylamino-17- (6-methoxy-) by flash chromatography (ethyl acetate, then 10% MeOH in ethyl acetate) Isoquinolin-1-yloxy) -2,14-dioxo-10-oxa-3,15-diaza-tricyclo [13.3.0.0 ^4,6 ] octadeca-7-ene-4-carboxylic acid ethyl ester was obtained as a white solid . LC-MS (Method A, retention time: 3.74 min), MS m / z 639 (M ⁺ +1).
Step 46c : Preparation of compounds 140 and 141:

(Z)-(1S, 4R, 13S, 17R) -13-tert-butoxycarbonylamino-17- (6-methoxy-isoquinolin-1-yloxy) -2,14-dioxo-10-oxa-3,15- Diaza-tricyclo [13.3.0.0 ^4,6 ] octadeca-7-ene-4-carboxylic acid ethyl ester was hydrolyzed according to Example 2 (Step 2E) to give Compound 140. Compound 140 is converted to Compound 141 according to Example 7, [(Z)-(1S, 4R, 13S, 17R) -4-cyclopropanesulfonylaminocarbonyl-17- (6-methoxy-isoquinolin-1-yloxy) -2, 14-Dioxo-10-oxa-3,15-diaza-tricyclo [13.3.0.0 ^4,6 ] octadeca-7-en-13-yl] -carbamic acid tert-butyl ester was converted. LC-MS (Method A, retention time: 3.51 min), MS m / z 714 (M ⁺ +1).
Example 47 : General procedure for the preparation of P4-carbamates from quaternary alcohols Preparation of compound 56: [(Z)-(1S, 4R, 14S, 18R) -4-cyclopropanesulfonylaminocarbonyl-18- (Isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-en-14-yl] -carbamic acid 1-methyl-cyclobutyl ester

工程47a：1-メチルシクロブタノールの製造：

Step 47a : Production of 1-methylcyclobutanol:

To a solution of 6 mL methylmagnesium iodide (3M in ether) at −10 ° C., cyclobutanone (1 g, 14.3 mmol) was added dropwise. The mixture was stirred for 2.5 hours (−10 ° C. to room temperature). Dilute HCl was added to the reaction mixture until all the precipitate was dissolved. The homogeneous reaction mixture was extracted with ether (2x). The combined ether extracts were dried (MgSO ₄ ), filtered and then concentrated under reduced pressure to give 0.9 g (73%) of 1-methylcyclobutanol as a yellow oil.
Step 47b . [(Z)-(1S, 4R, 14S, 18R) -4-cyclopropanesulfonylaminocarbonyl-18- (isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [ 14.3.0.0 ^4,6 ] Production of nonadeca-7-en-14-yl] -carbamic acid 1-methyl-cyclobutyl ester

To a slurry of KH (400 mg, 35% in oil, 3.5 mmol) was added 5 mL of hexane. It was stirred for 5 minutes and then hexane was removed. This process was repeated three times. THF (5 mL) was added to the prewashed KH, followed by 1-methyl-cyclobutanol (200 mg, 2.3 mmol) at 0 ° C. The mixture was stirred for 1 hour (0 ° C. to room temperature) and then carbonic acid dipyridin-2-yl ester (753 mg, 3.5 mmol) was added. The mixture was stirred at room temperature for 4 hours and then the reaction was diluted with sat. Aq. NH ₄ Cl. The mixture was then extracted with EtOAc and then the solvent was removed under reduced pressure to give 200 mg of crude carbonic acid 1-methyl-cyclobutyl ester pyridin-2-yl ester as an orange solid. It was used directly in the next step without further purification.

50 mg (0.073 mmol) of compound 11 [1- (2-amino-3,3-dimethyl-butyryl) -4- (6-methoxy-isoquinolin-1-yloxy) -pyrrolidine-2-carboxylic acid in CH ₂ Cl ₂ Suspension of acid (1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropyl) -amide bishydrochloride] with diisopropylethylamine (29 mg, 0.22 mmol) and 70 mg (0.33 mmol) of the above reagent (1-methyl carbonate- Cyclobutyl ester pyridin-2-yl ester) was added. The reaction mixture was stirred at room temperature overnight and then concentrated under reduced pressure. The residue was purified by preparative HPLC to give 32 mg (62%) of compound 56 as a white solid. Preparative HPLC conditions: YMC XTERRA, S5, 19x100mm, gradient: 50% B to 90% B, 15 min, retention 2 min, flow rate 25 mL / min. LC-MS (Method A, retention time: 3.47 min), MS m / z 708 (M ⁺ +1).

Compound 59 was prepared by the general method used for compound 56 (Example 47): 1-methylcyclopropanol was converted to carbonic acid 1-methyl-cyclopropyl ester pyridin-2-yl ester and the above method (Example 47) was prepared. ). 1-methyl-1-cyclopropanol was prepared by slightly modifying the method described in Kulinkovich, OG; Sviridov, SV; and Vasilevski, DA Synthesis, 1991, 234: methyl acetate (1.85 in diethyl ether (80 mL)). g, 25 mmol) and Ti (OPr-i) ₄ (744 μL, 2.5 mmol) to a stirred solution of ethyl magnesium bromide (3 M solution in diethyl ether) (18 mL, 53 mmol) in diethyl ether (60 mL). It was gradually added over 1 hour while maintaining at 18-20 ° C. Stirring is continued for 10 minutes when the addition is complete. The mixture was poured into cold (5 ° C.) 10% aq. H ₂ SO ₄ (250 mL) and the product was extracted with diethyl ether (3 × 50 mL). The mixed ether extract was washed with water (50 mL), dried (MgSO ₄ ) and then carefully concentrated by rotary evaporation (˜20 torr, rt) 1.7 g of 1-methyl-1-cyclopropanol (purity˜ 80%). ¹ H NMR (300 MHz, CDCl ₃ ) δ 0.42-0.45 (m, 2H), 0.73-0.77 (m, 2H), 1.44 (s, 3H), 1.64 (br s, 1H).

  The following compounds were also prepared using the general procedure described above: Compound 50 was prepared from 1-methylcyclopentanol; Compounds 39 and 51 were from 1,1,1-trifluoro-2-methylpropan-2-ol. Compound 55 was prepared from 1,1-difluoro-2-methylpropan-2-ol (1,1-difluoro-2-methylpropan-2-ol was described in Dickey, JB and Towne, EB US2700686. Compounds 40 and 52 were prepared from 1-chloro-2-methylpropan-2-ol, compound 124 was prepared from 1,1,1-trichloro-2-methylpropan-2-ol, Compound 123 was prepared from 3-hydroxy-3-methylbutan-2-one, and compound 139 was prepared from S-tert-butyl O-4-nitrophenylcarbonothioate (as described below) (this reagent was EM Gordon , JC Barrish, GS Bisacchi, CQ. Sun, JA Tino, GD Vite, and R. Zahler Produced as described in US005559256A).

実施例48：化合物126の製造

Example 48 : Preparation of compound 126

Compound 11 {cyclopropanesulfonic acid [14-amino-18- (isoquinolin-1-yloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca- in 2 mL DCM To a mixture of 7-ene-4-carbonyl] -amidobishydrochloric acid} (20 mg, 0.029 mmol) was added 18 μL (0.10 mmol) DIPEA and 4 mg (0.04 mmol) t-butyl isocyanate. The mixture was stirred overnight at room temperature. It was then diluted with EtOAc and then washed with pH 4 buffer (2x) and brine (1x). The organic layer was dried (MgSO ₄ ) and then concentrated under reduced pressure to give the crude product. The resulting oil was dissolved in methanol and purified by preparative HPLC (YMC XTERRA, S5, 19x100 mm, gradient: 50% B to 100% B, 15 min, retention 2 min, flow rate 25 mL / min) to give compound 126 white Obtained as a powder (17 mg, 82%): LC-MS (Method D, retention time: 3.24 min), MS m / z 695 (M ⁺ +1). ¹ H NMR (300 MHz, CDCl ₃ ) δ 0.86-0.96 (m, 2H), 1.12 (s, 9H), 1.01-1.49 (m, 18H), 1.52-1.66 (m, 1H), 1.70-1.78 (m, 1H), 1.83-1.95 (m, 2H), 2.14 -2.25 (m, 1H), 2.49 (brs, 1H), 2.71 (m, 2H), 2.86-2.92 (m, 1H), 4.07 (dd, J = 11.71, 4.03Hz, 1H), 4.40-4.44 ( m, 1H), 4.62-4.69 (m, 2H), 4.94-5.00 (m, 1H), 5.65-5.74 (m, 1H), 5.97 (br s, 1H), 7.03 (s, 1H), 7.32 (d , J = 6.22Hz, 1H), 7.50-7.52 (m, 1H), 7.68-7.78 (m, 2H), 8.00 (d, J = 5.86Hz, 1H), 8.22 (d, J = 7.68Hz, 1H) 10.15 (s, 1H).
Example 49 : Preparation of Compound 58:

Step 49a : 1-{[1- (2-tert-butoxycarbonylamino-non-8-enoyl) -4- (tert-butyl-dimethylsilanyloxy) -pyrrolidine-2-carbonyl] -amino} -2- Production of vinylcyclopropanecarboxylic acid ethyl ester

1-{[1- (2 (S) -tert-butoxycarbonylamino-nona-8-enoyl) -4 (R) -hydroxy-pyrrolidine-2 (S) carbonyl]-(1R)-in 10 mL DMF To a mixture of amino} -2 (S) -vinyl-cyclopropanecarboxylic acid ethyl ester (1.5 g, 2.87 mmoL), add imidazole (0.25 g, 3.67 mmoL) and tert-butyl-dimethylsilyl chloride (516 mg, 3.44 mmoL). It was. The mixture was stirred at room temperature for 2 days. The reaction mixture was then concentrated under reduced pressure and the residue was then dissolved in ethyl acetate. This solution was washed with water, dried over magnesium sulfate and then concentrated under reduced pressure to give a crude solid. Purify by flash chromatography (eluting with 20% ethyl acetate in hexane) to 1.43 g (78%) of 1-{[1- (2-tert-butoxycarbonylamino-non-8-enoyl) -4- (tert -Butyldimethylsilanyloxy) pyrrolidine-2-carbonyl] -amino} -2-vinylcyclopropanecarboxylic acid ethyl ester was obtained as a white solid.
¹ H NMR (300 MHz, CD ₃ OD) δ 0.10 (s, 6H), 0.89 (s, 9H), 1.22 (m, 3H), 1.31-1.48 (m, 16H), 1.50-1.75 (m, 3H), 2.06 (m, 3H), 2.11-2.33 (m, 2H), 3.70 (m, 2H), 4.03-4.19 (m, 2H), 4.21 (m, 1H), 4.45 (t, J = 7.87Hz, 1H) , 4.59 (m, 1H), 4.91 (d, J = 9.15Hz, 1H), 4.98 (d, J = 17.20Hz, 1H), 5.08 (dd, J = 10.25, 1.83Hz, 1H), 5.27 (dd, J = 17.38, 1.65 Hz, 1H), 5.65-5.87 (m, 2 H). LC-MS (Method A, retention time: 4.00 min), MS m / z 636 (M ⁺ +1).
Step 49b : 14-tert-butoxycarbonylamino-18- (tert-butyldimethyl-silanyloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-ene-4 -Production of carboxylic acid ethyl ester

1-{[1- (2-tert-Butoxycarbonylamino-non-8-enoyl) -4- (tert-butyl-dimethyl-silanyloxy) -pyrrolidine-2-carbonyl] -amino}-in 640 mL of methylene chloride To a solution of 2-vinyl-cyclopropanecarboxylic acid ethyl ester (1.63 g, 2.56 mmoL), 215 mg (0.26 mmoL) of tricyclohexylphosphine [1,3-bis (2,4,6-tri [benzylidene] ruthenium (IV The mixture was heated to reflux for 15 minutes and the residue was concentrated under reduced pressure and then purified by flash chromatography eluting with 30% ethyl acetate / hexanes.The sample was further decolorized and the crude product in hexanes. A second chromatography eluting with 50% ether of 1.5 g (96%) of 14-tert-butoxycarbonylamino-18- (tert-butyl-dimethyl-silanyloxy) -2,15-dioxo-3,16-diaza -Tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-ene-4-carboxylic acid The ethyl ester was obtained as a white solid: ¹ H NMR (500 MHz, CD ₃ Cl) δ 0.06 (s, 3H), 0.07 (s, 3H), 0.86 (s, 9H), 1.18-1.24 (m, 6H), 1.34-1.64 (m, 14H), 1.86-1.96 (m, 3H), 2.02-2.09 (m, 1H), 2.11-2.17 (m, 1H), 2.19-2.28 (m, 1H), 2.57-2.63 (m , 1H), 3.50-3.54 (m, 1H), 3.71 (dd, J = 10.22, 6.26Hz, 1H), 4.06-4.17 (m, 2H), 4.52-4.58 (m, 2H), 4.75 (d, J = 8.55Hz, 1H), 5.21 (t, J = 9.92Hz, 1H), 5.35 (d, J = 7.63Hz, 1H), 5.45-5.50 (m, 1H), 6.94 (s, 1H). LC-MS (Method A, retention time: 3.88 min), MS m / z 608 (M ⁺ +1).
Step 49c : 14-tert-butoxycarbonylamino-18- (tert-butyl-dimethyl-silanyloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-ene- 4-carboxylic acid production

14-tert-butoxycarbonylamino-18- (tert-butyldimethylsilanyloxy) -2,15-dioxo-3, in a mixed solvent system of THF (4 mL), methanol (1 mL), and water (2 mL), Powdered lithium hydroxide monohydrate (1.0 g, 50 mmoL) in a solution of 16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-ene-4-carboxylic acid ethyl ester (1.5 g, 2.47 mmoL) Was added. The pale yellow slurry was stirred at room temperature under N ₂ for 4 hours. The mixture was then concentrated under reduced pressure and the residue was partitioned between ether and water. The ether phase was discarded and the aqueous phase was treated with 1N HCl until pH 4 was reached. The acidic solution was extracted with EtOAc (3x). The combined EtOAc extracts were dried (MgSO ₄ ) and then concentrated under reduced pressure to give 1.2 g (84%) of 14-tert-butoxycarbonylamino-18- (tert-butyl-dimethyl-silanyloxy) -2,15- Dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-ene-4-carboxylic acid was obtained as an off-white solid. ¹ H NMR (300 MHz, CD ₃ OD) 0.12 (s, 6H), 0.89 (s, 9H), 1.23-1.64 (m, 17H), 1.70-1.87 (m, 1H), 1.90-2.49 (m, 6H) , 3.70-3.80 (m, 1H), 3.83-3.90 (m, 1H), 4.28-4.36 (m, 1H), 4.47-4.55 (m, 1H), 4.65 (s, 1H), 5.30-5.39 (m, 1H), 5.53-5.62 (m, 1H). LC-MS (Method A, retention time: 3.69 min), MS m / z 580 (M ⁺ +1).
Step 49d : [18- (tert-Butyl-dimethyl-silanyloxy) -4-cyclopropanesulfonylaminocarbonyl-2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-ene Of -14-yl] -carbamic acid tert-butyl ester

14-tert-Butoxycarbonylamino-18- (tert-butyldimethylsilanyloxy) -2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-ene-4-carvone acid (500 mg, 0.86 mmol) was dissolved in THF and 25 mL, CDI (180 mg, 1.12 mmol) was treated with (using an oven dried glass vessel was careful to avoid moisture by keeping it dry N ₂ atmosphere). After the reaction mixture was refluxed for 2 hours, it was cooled to room temperature and then treated sequentially with cyclopropylsulfonamide (135 mg, 1.12 mmoL) and DBU (170 mg, 1.12 mmoL). The reaction mixture was stirred at room temperature for 4 hours and then the THF was removed by rotary evaporation. The residue was partitioned between ethyl acetate and pH 4 buffer. The organic layer was dried (MgSO ₄ ) and then concentrated under reduced pressure to give the crude product. It was then purified by flash chromatography (eluting with 33% ethyl acetate in hexane) and 300 mg (51%) of [18- (tert-butyl-dimethyl-silanyloxy) -4-cyclopropanesulfonylaminocarbonyl-2, 15-Dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-en-14-yl] -carbamic acid tert-butyl ester was obtained as a white solid. ¹ H NMR (300 MHz, CD ₃ OD) δ 1H 0.07 (s, 3H), 0.08 (s, 3H), 0.85 (s, 9H), 0.87-1.49 (m, 21H), 1.73-1.95 (m, 3H) 2.08-2.16 (m, 1H), 2.25-2.36 (m, 2H), 2.42-2.56 (m, 1H), 2.85-2.93 (m, 1H), 3.65-3.74 (dd, J = 10.61, 3.66Hz, 1H), 3.89 (d, J = 10.25Hz, 1H), 4.34 (m, J = 9.70, 9.70Hz, 1H), 4.43 (t, J = 7.87Hz, 1H), 4.57 (s, 1H), 4.94- 5.01 (m, 1H), 5.10 (d, J = 8.78Hz, 1H), 5.66-5.75 (m, 1H), 6.55 (s, 1H), 10.13 (s, 1H). LC-MS (Method A, retention Time: 3.81 min), MS m / z 683 (M ⁺ +1).
Step 49e : (4-Cyclopropanesulfonylaminocarbonyl-18-hydroxy-2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-en-14-yl) -carbamic acid tert-butyl ester

25mLのTHF中の[18-(tert-ブチル-ジメチルシラニルオキシ)-4-シクロプロパンスルホニル-アミノカルボニル-2,15-ジオキソ-3,16-ジアザ-トリシクロ[14.3.0.0^4,6]ノナデカ-7-エン-14-イル]-カルバミン酸tert-ブチルエステル(330mg、0.48mmoL)の混合物にテトラブチルアンモニウムフロリド (150mg、0.54mmoL)を加えた。反応混合物を室温で18時間撹拌し、次いでTHFをロータリーエバポレーションで除去した。残渣を酢酸エチルと水の間に分配した。有機層を乾燥し(MgSO₄)、次いで減圧下で濃縮して粗生成物を得た。次に、それをヘキサンでトリチュレートして200mg(73%)の(4-シクロプロパンスルホニルアミノカルボニル-18-ヒドロキシ-2,15-ジオキソ-3,16-ジアザ-トリシクロ[14.3.0.0^4,6]ノナデカ-7-エン-14-イル)-カルバミン酸tert-ブチルエステルを白色固体として得た。¹H NMR (500MHz、CD₃Cl) δ 1.87-1.64 (m、21H)、1.70-1.98 (m、3H)、2.15-2.56 (m、5H)、2.85-2.94 (m、1H)、3.71 (d、J=13.91Hz、1H)、4.10-4.26 (m、2H)、4.51 (t、J=7.87Hz、1H)、4.62 (s、1H)、4.98 (m、1H)、5.06 (d、J=8.78Hz、1H)、5.64-5.71 (m、1H)、6.72 (s、1H)、10.24 (s、1H). LC-MS (方法A、保持時間：2.85min)、MS m/z 569 (M⁺+1)。
工程49f：化合物58：[(Z)-(1S,4R,14S,18R)-4-シクロプロパンスルホニルアミノカルボニル-2,15-ジオキソ-18-(3-フェニル-イソキノリン-1-イルオキシ)-3,16-ジアザ-トリシクロ[14.3.0.0^4,6]ノナデカ-7-エン-14-イル]-カルバミン酸tert-ブチルエステルの製造

[18- (tert-Butyl-dimethylsilanyloxy) -4-cyclopropanesulfonyl-aminocarbonyl-2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca in 25 mL THF To a mixture of -7-en-14-yl] -carbamic acid tert-butyl ester (330 mg, 0.48 mmoL) was added tetrabutylammonium fluoride (150 mg, 0.54 mmoL). The reaction mixture was stirred at room temperature for 18 hours and then the THF was removed by rotary evaporation. The residue was partitioned between ethyl acetate and water. The organic layer was dried (MgSO ₄ ) and then concentrated under reduced pressure to give the crude product. It was then triturated with hexane to give 200 mg (73%) of (4-cyclopropanesulfonylaminocarbonyl-18-hydroxy-2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] Nonadeca-7-en-14-yl) -carbamic acid tert-butyl ester was obtained as a white solid. ¹ H NMR (500 MHz, CD ₃ Cl) δ 1.87-1.64 (m, 21H), 1.70-1.98 (m, 3H), 2.15-2.56 (m, 5H), 2.85-2.94 (m, 1H), 3.71 (d , J = 13.91Hz, 1H), 4.10-4.26 (m, 2H), 4.51 (t, J = 7.87Hz, 1H), 4.62 (s, 1H), 4.98 (m, 1H), 5.06 (d, J = 8.78Hz, 1H), 5.64-5.71 (m, 1H), 6.72 (s, 1H), 10.24 (s, 1H). LC-MS (Method A, retention time: 2.85min), MS m / z 569 (M ⁺ +1).
Step 49f : Compound 58: [(Z)-(1S, 4R, 14S, 18R) -4-cyclopropanesulfonylaminocarbonyl-2,15-dioxo-18- (3-phenyl-isoquinolin-1-yloxy) -3 Of 1,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-en-14-yl] -carbamic acid tert-butyl ester

(4-Cyclopropanesulfonylaminocarbonyl-18-hydroxy-2,15-dioxo-3,16-diaza-tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-en-14-yl)-in 1 mL DMF- To a mixture of carbamic acid tert-butyl ester (20 mg, 0.035 mmol) potassium tert-butoxide (22 mg, 0.196 mmol) was added. The mixture was stirred at room temperature for 5 minutes and then 1-chloro-3-phenylisoquinoline (15 mg, 0.062 mmol) was added. The reaction mixture was stirred at room temperature for 15 hours and then concentrated under reduced pressure. The crude product was triturated with ether. The residue was dissolved in MeOH and then purified by preparative HPLC (YMC XTERRA, S5, 19x100 mm, gradient: 60% B-100% B, 15 min, retention 2 min, flow rate 25 mL / min) to give 10 mg (39%) of compound 58, [(Z)-(1S, 4R, 14S, 18R) -4-cyclopropanesulfonylaminocarbonyl-2,15-dioxo-18- (3-phenyl-isoquinolin-1-yloxy) -3,16-diaza -Tricyclo [14.3.0.0 ^4,6 ] nonadeca-7-en-14-yl] -carbamic acid tert-butyl ester was obtained as a white solid. ¹ H NMR (500 MHz, CD ₃ Cl) δ 0.90-1.64 (m, 21H), 1.76-1.97 (m, 3H), 2.26-2.31 (m, 1H), 2.52-2.63 (br s, 1H), 2.69- 2.81 (m, 2H), 2.88-2.93 (m, 1H), 4.11 (d, J = 11.60Hz, 1H), 4.33 (m, 1H), 4.61 (d, J = 7.94Hz, 2H), 4.99 (t , J = 9.31Hz, 1H), 5.05 (d, J = 7.93Hz, 1H), 5.69-5.74 (m, 1H), 6.08 (s, 1H), 6.60 (s, 1H), 7.38-7.47 (m, 2H), 7.50 (t, J = 7.63Hz, 2H), 7.63 (t, J = 7.32Hz, 1H), 7.71 (s, 1H), 7.77 (d, J = 8.24Hz, 1H), 8.10 (d, . J = 7.32Hz, 2H), 8.18 (d, J = 7.93Hz, 1H), 10.27 (s, 1H) LC-MS ( method A, retention time: 3.72min), MS m / z 772 (M + + 1).

  The compounds in Tables 2 and 3 were prepared using the methods described in the above examples.

表2

Table 2

表3

Table 3

  The compounds in Tables 4 and 5 could be prepared using the methods described in the above examples.

表4

Table 4

表5

Table 5

Example 50
Biological test
Recombinant HCV NS3 / 4A protease complex FRET peptide assay

  The purpose of this in vitro assay was to measure the inhibition of the HCV NS3 protease complex from the following BMS strain, H77C strain, or J4l6S strain by the compounds of the present invention. This assay provides an indication of how effective the compounds of the invention are in inhibiting HCV proteolytic activity.

  Serum from HCV-infected patients was obtained from Dr. T. Wright, San Francisco Hospital. Manipulated full-length cDNA (complementary deoxyribonucleic acid) template of HCV genome (BMS strain) was selected based on homology between other genotype 1a strains by reverse transcription PCR (RT-PCR) of serum RNA (ribonucleic acid) Obtained using primers. From determination of the complete genome sequence, genotype 1a was assigned to an HCV isolate according to the classification of Simmonds et al. (See P Simmonds, KA Rose, S Graham, SW Chan, F McOmish, BC Dow, EA Follett, PL Yap, and H Marsden, J. Clin. Microbiol., 31 (6), 1493-1503 (1993)). The amino acid sequence of the nonstructural region, NS2-5B, was shown to be> 97% identical to HCV genotype 1a (H77C) and 87% identical to genotype 1b (J4L6S). Infectious clones, H77C (1a genotype) and J4L6S (1b genotype) were obtained from R. Purcell (NIH), which sequences are Genbank (AAB67036, Yanagi, M., Purcell, RH, Emerson, SU and Bukh, J Proc. Natl. Acad. Sci. USA 94 (16), 8738-8743 (1997); AF054247, Yanagi, M., St Claire, M., Shapiro, M., Emerson, SU, Purcell, RH and Bukh , J, Virology 244 (1), 161-172. (1998)).

Recombinant NS3 / 4A protease complex was produced using H77C and J4L6S strains. DNA encoding the recombinant HCV NS3 / 4A protease complex (amino acids 1027 to 1711) of these strains was engineered as described in P. Gallinari et al. (Gallinari P, Paolini C, Brennan D, Nardi C, Steinkuhler C, De Francesco R. Biochemistry. 38 (17): 5620-32, (1999)). Briefly, a 3 lysine solubilized tail was added to the 3 ′ end of the NS4A coding region. In order to avoid proteolytic cleavage of lysine tag, cysteine (amino acid 1711) in the P1 position of the NS4A-NS4B cleavage site was converted to glycine. In addition, a cysteine to serine mutation was introduced by PCR at amino acid position 1454 to avoid autolytic cleavage of the NS3 helicase domain. Protocol described in P. Gallinari et al. (See Gallinari P, Brennan D, Nardi C, Brunetti M, Tomei L, Steinkuhler C, De Francesco R., J Virol. 72 (8): 6758-69 (1998)). According to (partially modified), the mutant DNA fragment was cloned into the pET21b bacterial expression vector (Novagen) and the NS3 / 4A complex was expressed in Escherichia coli strain BL21 (DE3) (Invitrogen). Briefly, NS3 / 4A expression was induced with 0.5 mM isopropyl β-D-1-thiogalactopyranoside (IPTG) at 20 ° C. for 22 hours. A typical fermentation (10 L) yielded approximately 80 g of wet cell paste. Cells were treated with 25 mM N- (2-hydroxyethyl) piperazine-N '-(2-ethanesulfonic acid) (HEPES), pH 7.5, 20% glycerol, 500 mM sodium chloride (NaCl), 0.5% Triton-X100, 1 micron Gram / milliliter (`` μg / mL '') lysozyme, 5 mM magnesium chloride (MgCl ₂ ), 1 ug / ml Dnase I, 5 mM β-mercaptoenanol (βME), protease inhibitor-ethylenediaminetetraacetic acid (EDTA) free ( Roche) was resuspended in lysis buffer (10 mL / g), homogenized and incubated at 4 ° C. for 20 minutes. The homogenate was sonicated and then ultracentrifuged at 235000 g for 1 hour at 4 ° C. to remove impurities. Imidazole was added to the supernatant to a final concentration of 15 mM, and the pH was adjusted to 8.0. Nickel-nitrilotriacetic acid (Ni-NTA) pre-equilibrated with buffer B (25 mM HEPES, pH 8.0, 20% glycerol, 500 mM NaCl, 0.5% Triton-X100, 15 mM imidazole, 5 mM βME) Loaded on the column. The sample was loaded at a flow rate of 1 mL / min. The column was washed with 15 column volumes of Buffer C (same as Buffer B except containing 0.2% Triton-X100). The protein was eluted with 5 column volumes of Buffer D (same as Buffer C except containing 200 mM imidazole).

  NS3 / 4A protease complex-containing fractions were pooled and pre-equilibrated with buffer D (25 mM HEPES, pH 7.5, 20% glycerol, 300 mM NaCl, 0.2% Triton-X100, 10 mM βME) Superdex- Loaded on S200. The sample was loaded at a flow rate of 1 mL / min. NS3 / 4A protease complex containing fractions were pooled and concentrated to approximately 0.5 mg / ml. The purity of NS3 / 4A protease complex from BMS, H77C and J4L6S strains was judged to be 90% or more by SDS-PAGE and mass spectrometry.

  The enzyme was stored at −80 ° C., thawed on ice, and diluted with assay buffer before use. The substrate used for NS3 / 4A protease assay is RET S1 (Resonance Energy Transfer Depsipeptide Substrate) described in Taliani et al., Anal. Biochem. 240 (2): 60-67 (1996); Inc. cat # 22991) (FRET peptide). The sequence of this peptide is largely based on the NS4A / NS4B natural cleavage site except that there is an ester bond rather than an amide bond at the cleavage site. The peptide substrate was incubated with one of the three recombinant NS3 / 4A in the presence or absence of the compounds of the invention, and the formation of the fluorescent reaction product was followed in real time using the Cytofluor Series 4000.

  The reagents are as follows. HEPES and glycerol (Ultrapure) were obtained from GIBCO-BRL. Dimethyl sulfoxide (DMSO) was obtained from Sigma. β-mercaptoenanol was obtained from Bio Rad.

  Assay buffer: 50 mM HEPES, pH 7.5; 0.15 M NaCl; 0.1% Triton; 15% glycerol; 10 mM βME. Substrate: 2 μM final concentration (from 2 mM stock solution in DMSO stored at −20 ° C.). HCV NS3 / 4A type 1a (1b), 2-3 nM final concentration (from 5 μM stock solution in 25 mM HEPES, pH 7.5, 20% glycerol, 300 mM NaCl, 0.2% Triton-X100, 10 mM βME). For compounds with potency near the assay limit, the assay was made more sensitive by adding 50 μg / ml bovine serum albumin (Sigma) to the assay buffer and reducing the final protease concentration to 300 pM.

  The assay was performed in a 96-well polystyrene black plate from Falcon. Each well contained 25 microliters (“μL”) of NS3 / 4A protease complex in assay buffer, 50 μL of the compound of the invention in 10% DMSO / assay buffer, and 25 μL of substrate in assay buffer. A control (no compound) was also prepared on the same assay plate. The enzyme complex was mixed with the compound or control solution 1 minute before adding the substrate and starting the enzyme reaction. The assay plate was read rapidly using a Cytofluor Series 4000 (Perspective Biosystems). The instrument was set to read 340 nm emission and 490 nm excitation at 25 ° C. Readings were usually taken for about 15 minutes.

The percent inhibition was calculated by the following formula:
100 − [(δF _inh / δF _con ) x100]
[Where δF is the change in fluorescence over the linear range of the curve. Apply nonlinear curve fit to inhibition-concentration data and use 50% effective concentration (IC ₅₀ ) as equation, y = A + ((BA) / (1 + ((C / x) ^ D))) and Excel X1 Calculated using fit software. ].

All test compounds were found to have an IC ₅₀ of 1.2 μM or less. Furthermore, the compounds of the present invention tested against two or more types of NS3 / 4A complexes have similar inhibitory properties, but the compounds are uniformly more potent against strain 1b than strain 1a. I understood.
Specificity assay

  Specificity assays were performed to show the sensitivity of the compounds of the invention in inhibiting HCV NS3 / 4A protease compared to other serine or cysteine proteases.

  The specificity of the compounds of the invention has been detected against various serine proteases: human neutrophil elastase (HNE), porcine pancreatic elastase (PPE), and human pancreatic chymotrypsin, and certain cysteine proteases: human liver cathepsin B. It was. In all cases, a 96-well plate format protocol using a colorimetric p-nitroaniline (pNA) substrate specific for each enzyme was used as described above (with some modifications for the serine protease assay) ( PCT patent application No. WO 00/09543). All enzymes were purchased from Sigma and substrates were purchased from Bachem.

  Each assay included a 2 hour enzyme-inhibitor preincubation at room temperature followed by ˜30% conversion (measured using a Spectramax Pro microplate reader) by substrate addition and hydrolysis. The compound concentration varied from 100 to 0.4 micromolar (“μM”) depending on its potency.

  The final concentration for each assay is as follows.

50 mmol (`` mM '') tris (hydroxymethyl) aminomethane / hydrochloric acid (Tris-HCl) pH 8, 0.5 M sodium sulfate (Na ₂ SO ₄ ), 50 mM NaCl, 0.1 mM EDTA, 3% DMSO, 0.01% Tween-20 :
Contains 133 μM succ-AAA-pNA and 20 nM HNE or 8 nM PPE; 100 μM succ-AAPF-pNA and 250 pM chymotrypsin.

100 mM NaHPO ₄ (Sodium hydrogen phosphate) pH 6, 0.1 mM EDTA, 3% DMSO, 1 mM TCEP (Tris (2-carboxyethyl) phosphine / hydrochloric acid), 0.01% Tween-20, 30 μM Z-FR-pNA and 5 nM cathepsin B (Enzyme stock was activated with 20 mM TCEP containing buffer prior to use.)

The percentage inhibition was calculated using the following formula:
[1-((UV _inh -UV _blank ) / (UV _ctl -UV _blank ))] x100

A nonlinear curve fit was applied to the inhibition-concentration data and a 50% effective concentration (IC ₅₀ ) was calculated using Excel X1 fit software.

The HCV reprion assay was utilized in the present invention and was prepared, performed and confirmed as follows.
HCV replicon generation

The HCV replicon whole cell line was established as described in Lohmann V, Korner F, Koch J, Herian U, Theilmann L, Bartenschlager R., Science 285 (5424): 110-3 (1999). This system allowed the effect of the HCV protease compounds of the present invention on HCV RNA replication. Briefly, HCV cDNA was synthesized by Operon Technologies, Inc. (Alameda, Calif.) Using the HCV strain 1B sequence described in Lohmann's paper (accession number: AJ238799), and then the full-length replicon was standard molecular biology. Was assembled into plasmid pGem9zf (+) (Promega, Madison, WI) using standard techniques. The replicon is (i) an HCV 5'UTR fused to the first 12 amino acids of the capsid protein, (ii) the neomachine phosphotransferase gene (neo), (iii) an IRP from encephalomyocarditis virus (EMCV), and (iv) It consists of HCV NS3-NS5B gene and HCV 3'UTR. Plasmid DNA was linearized with ScaI and then RNA transcripts were synthesized in vitro using the T7 MegaScript transcription kit (Ambion, Austin, TX) according to the manufacturer's instructions. 10 micrograms (“μg”) RNA transcript of 4x10 ⁶ Huh-7 cells (provided by R. Bartenschlager, available from Health Science Research Resources Bank, Japan Health Sciences Foundation) to create cell lines The material was electroporated (GenePulser System, Bio-Rad) and plated into 100 mm dishes. After 24 hours, selective medium G418 containing 1.0 milligram / milliliter (“mg / ml”) was added and the medium was changed every 3-5 days. Approximately 4 weeks after electroporation, a small colony was seen, which was isolated and expanded for further analysis. These cell lines were treated with 10% heat-inactivated calf serum (Sigma), 10 mL 100X penicillin / streptomycin (Cat # 15140-122) Gibco-BRL, Rockville, MD, 1 mg / ml Geneticin (Cat # 10131-027) DMEM containing Gibco-BRL, Rockville, MD (Cat # 11965-084) Maintained in Gibco-BRL, Rockville, MD at 37 ° C., 5% CO ₂ , 100% relative humidity. One of the cell lines with approximately 3,000 copies of HCV replicon RNA / cell (deposited with American Type Culture Collection under ATCC Deposit No. PTA-4583) was used for assay development (HCV 1b-377-neo replicon cells) .
FRET assay

Huh7 cells constitutively expressing the HCV replicon were grown in Dulbecco's Modified Eagle Medium (DMEM) containing 10% fetal calf serum (FCS) and 1 mg / ml G418 (Gibco-BRL). Cells were seeded the night before in 96-well tissue culture sterile plates (1.5 × 10 ⁴ cells / well). Compound and no compound controls were prepared in dilution plates using DMEM containing 4% FCS, 1: 100 penicillin / streptomycin, 1: 100 L-glutamine and 5% DMSO (final concentration in assay 0.5% DMSO). The compound / DMSO mixture was added to the cells and incubated at 37 ° C. for 4 days. After 4 days, cells were first assessed for cytotoxicity using Alamar Blue (Trek Diagnotstic Systems) to read CC50. Compound toxicity (CC50) was measured by adding 1/10 volume of Alamar Blue to the medium incubating the cells. After 4 hours, the fluorescence signal from each well was read using a Cytofluor Series 4000 (Perspective Biosystems) for excitation wavelength at 530 nm and emission wavelength at 580 nm. The plates were then rinsed thoroughly with phosphate buffered saline (PBS) (3 times, 150 μL). HCV protease substrate-containing lysis assay reagent (5X cell luciferase cell culture lysis reagent diluted 1X with distilled water (Promega # E153A) NaCl added to final 150 mM, and FRET diluted from 2 mM stock in 100% DMSO to final 10 μM Cells were lysed with 25 μL of peptide). An HCV protease substrate (FRET peptide; described in AnaSpec, Inc. cat # 22991, Taliani et al., Anal. Biochem. 240 (2): 60-67 (1996)) is a fluorescent donor, EDANS, near one end of the peptide. , And other acceptors near the end, DABCYL. Although the fluorescence of the peptide is attenuated by intermolecular resonance energy transfer (RET) between the donor and acceptor, NS3 protease cleaves the peptide, so the product is released from the RET decay, revealing the fluorescence of the donor. The plate was then placed in a Cytofluor 4000 instrument set to 340 nm excitation / 490 nm emission, 21-cycle automatic mode, and the plate was read in kinetic mode at 25 ° C. The reaction was usually followed for about 15 minutes.

The percent inhibition of both effect and toxicity was calculated using the following equation:
100 − [(δF _inh / δF _con ) x100]
[Where δF is the change in fluorescence over the linear range of the curve. Apply nonlinear curve fit to inhibition-concentration data, using 50% effective concentration (IC ₅₀ and CC ₅₀ ) equation, y = A + ((BA) / (1 + ((C / x) ^ D))) Calculated using Excel X1 fitting software. ].
Luciferase assay

As a secondary assay, EC ₅₀ measurements from the replicon FRET assay were confirmed using a luciferase reporter assay. The use of the replicon luciferase reporter assay was first described by Krieger et al (Krieger N, Lohmann V, and Bartenschlager R, J. Virol. 75 (10): 4614-4624 (2001)). The replicon construct that describes our FRET assay consists of a cassette containing a Renilla Luciferase gene fused with a neomycin resistance gene with a sequence representing the NS3 cleavage site (NS4A / B site), followed by a Blasicidin resistance gene. It was modified by replacing with the restriction site Asc1 / Pme1 used for subcloning. An adaptive mutation at position 1179 (serine to isoleucine) was also introduced (Blight KJ, Kolykhalov, AA, Rice, CM, Science 290 (5498): 1972-1974). A cell line containing the replicon was selected and maintained in 1.25 μg / ml Blasticidin. The luciferase reporter assay was prepared the night before by seeding replicon cells at a density of 7000 cells / well in a 96 well plate. After 1 day, the medium was replaced with fresh DMEM containing 4% FBS and compound dilutions were prepared and added to a final concentration of 0.5% DMSO (final medium volume 150 μl). After further incubation for 4 days in a 37 ° C./5% CO ₂ incubator, the Renilla luciferase activity was analyzed using the Dual-Glo luciferase assay system. Medium (100 μl) was removed from each well. To 50 μl of the remaining medium, 50 μl of Dual-Glo luciferase reagent was added and then the plate was shaken at room temperature for an additional 10 minutes to 2 hours. Next, Dual-Glo Stop & Glo reagent (50 μl) was added to each well and the plate was again shaken at room temperature for 10 minutes to 2 hours. Plates were read using a Packard TopCount NXT using a luminescence program.

The percentage inhibition was calculated using the following formula:

Values were graphed and analyzed using XL fit to obtain EC ₅₀ values.

Representative compounds of the invention were evaluated using several of the HCV NS3 / 4A protease recombinase assay, HCV replicon cell-based assay and / or outlined specificity assay. For example, Compound 2 was found to have an IC50 of 26 nM for BMS NS3 / 4A strain protease in the enzyme assay. Similar potency values were obtained using the published H77C (IC ₅₀ , 4.2 nM) and J4L6S (IC ₅₀ , 1.9 nM) strains. The EC ₅₀ value in the replicon assay was 146 nM.

  In the specificity assay, the compounds were found to have the following activities: HNE> 100 μM; PPE> 100 μM; chymotrypsin> 100 μM; cathepsin B> 100 μM. These results indicate that this family of compounds is highly specific for NS3 protease, and many of these members inhibit HCV replicon replication.

The test compound was found to have the activity in the following range.
IC ₅₀ activity range (NS3 / 4A BMS strain): A <50 μM; B <5 μM; C <0.5 μM; D <0.05 μM.
EC ₅₀ activity range (for test compounds): A <50 μM; B <5 μM; C <0.5 μM; D <0.05 μM.
Tables 6 & 7: Compound activity table

表6

Table 6

表7

Table 7

Combination test

MS(ESI) m/z= 691.2 (MH⁺)；HPLC rt 1.21min；純度(99%)、およびNS5Bレプリカーゼ阻害剤(化合物2006；WO 03/010141)は大環状HCV NS3プロテアーゼ阻害剤、化合物28を用いる2剤の組み合わせにおいて試験した。さらに、化合物28は、NS5AおよびNS5B阻害剤との3剤の組み合わせについても試験した。 Because clinical drug resistance is often manifested in viral infection after single drug therapy, it is necessary to evaluate the additive, antagonistic, or synergistic properties of combination therapy. As described above, the present inventors evaluated the availability of the NS3 protease macrocyclic inhibitor of the present invention in combination therapy using an inhibitor targeting Intron A and other HCV proteins using the HCV replicon system as described above. did. Three HCV antiviral drugs, Intron A (recombinant interferon α-2b), HCV NS5A inhibitor with the following structure:

MS (ESI) m / z = 691.2 (MH ⁺ ); HPLC rt 1.21 min; purity (99%), and NS5B replicase inhibitor (compound 2006; WO 03/010141) is a macrocyclic HCV NS3 protease inhibitor, compound 28 Tested in a two-drug combination using In addition, compound 28 was also tested for a triple combination with NS5A and NS5B inhibitors.

For the experiments shown in Tables 8 and 9, inhibitors were tested at 11 concentrations each. 200x stock solutions of each inhibitor concentration were prepared by diluting 2 or 3 times with DMSO, then cells / medium was added. The drugs were tested in monotherapy and in combination with NS3 protease inhibitors at various concentration ratios. Cells were exposed to the compound for 4 days and then the amount of HCV inhibition was measured using the FRET assay. The possible cytotoxicity of these combined drugs was analyzed in parallel by Alamar Blue staining. The CC ₅₀ value of the indicated compound was greater than the highest concentration of the test inhibitor. The degree of antagonism, additivity (additive action), or synergy (synergism) was measured over a range of drug concentrations, and a combination response curve was applied to drug treatment to assess the antiviral effect of the drug treatment combination. . Concentration ratios were analyzed using the Chou method (Chou, Ting-Chao, and Rideout (editor), (1991) Synergism and Antagonism in Chemotherapy, P. 61-101, Academic Press, New York). Tables 8 and 9 report the estimated EC ₅₀ values and combination index (CI) of the test compounds. All estimates were calculated using SAS Proc NLIN and a two parameter logistic. All combination indices were tested for deviation from additivity using the isobologram method. Asymptotic confidence intervals were also calculated for each combination index. These intervals were used to test deviations from additivity by comparing the boundaries to some-indicating antagonism when the lower limit of the interval is greater than 1 and synergism when the upper limit is less than 1. The value of 1 included in the interval indicates additivity.

In addition to the above combination index approach, Universal Response Surface Approach (URSA) was used to evaluate the antiviral effect of a combination of NS5A or NS5B inhibitor and NS3 protease inhibitor. The experiment was performed using a matrixed format of 8 concentrations of NS3 protease inhibitor that crossed against 10 concentrations of NS5A or NS5B inhibitor in each of 3 plates. 200x stock solutions at each inhibitor concentration were prepared by 3-fold dilution in DMSO. The effect on replication was evaluated using the HCV replicon system as described above. Data were analyzed using URSA as described in Greco, Park and Rustum (Greco, Park, Sook, and Rustum (1990) Application of a New Approach for the Quantitation of Drug Synergism to the Combination of cis-Diamminedichloplatinum and 1- β-D-Arabinofuranosylcytosine, Cancer Research, 50, 5318-5327). The interaction of the two drugs was evaluated using non-linear regression using a bisection method. Seven parameters were evaluated. These result in a minimum response or no drug response, a maximum response or infinite drug response, two drug EC _{50 s} , two drug slope parameters, and synergy, antagonism, or assessment of additivity Contains the drug interaction parameter α. Tables 10 and 11 show key estimation parameters and standard errors. When α is equal to 0, it means additiveity, when the interaction parameter is greater than 0, it means synergy, and when the interaction parameter is less than 0, it means antagonism. Also shown is the confidence interval for the interaction parameter. Use this interval to test the deviation from additivity by comparing the boundary with 0-an interval that shows synergy when the lower limit of the interval is greater than 1 and antagonism when the upper limit is less than 1 A value of 1 included in indicates an additivity.

Table 8 summarizes the data obtained from the combination of Compound 28 and Intron A. Also shown are EC ₅₀ values for each monotherapy. In two experiments, the combination of Compound 28 and Intron A produced either synergism (synergy) or additive action (synergy) at 75%, 90, and 95% effective doses.

  The combined effects of Compound 28 with HCV NS5A inhibitors are summarized in Table 9. In two experiments, synergistic or additive effects were seen at 75, 90, and 95% effective doses of Compound 28. Importantly, there was no drug antagonism at 75, 90, or 95% effective dose when Compound 28 was combined with an HCV NS5A inhibitor or interferon alpha-2b. Validation experiments with these same inhibitor combinations in matrix format (Table 10) yielded results similar to the combination index experiments (additive with the combination of NS5A inhibitor and compound 28).

  The matrix format was also used to test the activity of the HCV NS5B inhibitor and compound 28 combination (Table 11). Overall, additivity was observed in two experiments.

  Compound 28 was also tested in a triple drug combination experiment with NS5A inhibitor and NS5B inhibitor (Table 12). The starting concentration of the monotherapy consisted of a 0.667 μM solution of NS5A inhibitor, a 0.3 μM solution of compound 28, and a 2.5 μM solution of NS5B inhibitor. The starting concentration of the triple combination contained a mixture of 1/3 of each of the above concentrations. Dose response curves for all 3 monotherapy and triple combination therapy were obtained using 3 fold dilutions and data analysis was performed as described above for the combination index experiment. A synergistic effect was observed using the triple combination. No drug antagonism was observed at 75, 90, or 95% effective dose with the three drug combinations.

These results indicate that combined treatment of replicon cells with inhibitors targeting HCV NS3 protease inhibitor, compound 28 and Intron A, and / or HCV NS5A and / or NS5B has an additive to synergistic antiviral effect. Arise. The ability to use these NS3 protease inhibitors in combination therapy can provide significant advantages over monotherapy in the treatment of HCV.
Table 8: Combination of two drugs using interferon alpha-2b

表9：NS5A阻害剤を用いる2薬剤の組み合わせ

Table 9: Combination of two drugs using NS5A inhibitor

表10：NS5A阻害剤を用いる2薬剤の組み合わせ：マトリックス試験

Table 10: Combination of two drugs with NS5A inhibitor: matrix test

表11：NS5B阻害剤を用いる2薬剤の組み合わせ：マトリックス試験

Table 11: Combination of two drugs with NS5B inhibitor: matrix study

表12：3薬剤の組み合わせ試験

Table 12: Combination test of 3 drugs

  Those skilled in the art will recognize that while the invention has been described above with reference to specific aspects, other aspects are intended to be within the scope of the claims. All documents mentioned in this specification constitute part of this specification.

Claims (56)

Formula I:

Or a pharmaceutically acceptable enantiomer, diastereomer, salt, solvate, or prodrug thereof [wherein
(a) R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , and R ₆ are each independently H; C _1-6 alkyl; C _3-7 cycloalkyl; C _1-6 alkoxy; C _{3 -7} cycloalkoxy; halo-C _1-6 alkoxy; halo-C _1-6 alkyl; cyano; halo; hydroxyl; C _1-6 alkanoyl; nitro; amino; mono- or di- (C _1-6 ) alkylamine; Mono or di- (C _3-7 ) cycloalkylamine; mono or di-C _1-6 alkylamide; mono or di- (C _3-7 ) cycloalkylamide; carboxyl; (C _1-6 ) carboxyester; thiol; C _1-6 thioalkyl; C _1-6 alkyl sulfoxide; C _1-6 alkyl sulfonate; C _1-6 alkyl sulfonamide; sometimes substituted with Het C _6-10 aryl; C _{7 - 14} alkyl Aryl; C _6-10 aryloxy; C _7-14 alkylaryloxy; 4-7 membered monocyclic heteroaryloxy; R _{1 to} R ₆ may be linked to an isoquinoline group via a C _1-6 alkyl linking group;
(b) R ₇ is NH ₂ or —NR ₁₀ R ₁₁ ; where R ₁₀ is C _1-6 alkyl, C _1-6 haloalkyl, C (O) —NR ₁₂ R ₁₃ , C (O) -OR ₁₄ , C (O) -SR ₁₅ , or -C (O) -R ₁₆ ; R ₁₁ is H, C _1-6 alkyl, or C _1-6 haloalkyl; provided that R ₁₂ or R ₁₁ is H when any of R ₁₃ is H;
R ₁₂ and R ₁₃ are each independently H; C _1-6 alkyl, C _3-7 cycloalkyl, or C _4-10 alkyl cycloalkyl (halo, C _1-3 alkoxy, C _1-3 halo, respectively) May be substituted with alkoxy, C _1-3 alkyl, or C _1-3 haloalkyl); or aryl; where R ₁₂ and R ₁₃ are taken together with the nitrogen to which they are attached. Can form a 4-7 membered heterocycle;
R ₁₄ and R ₁₅ are each independently C _1-6 alkyl, C _3-7 cycloalkyl, or C _4-10 alkyl cycloalkyl (halo, C _1-3 alkoxy, C _1-3 haloalkoxy, C May be substituted with _1-3 alkyl, or C _1-3 haloalkyl); aryl, or Het; R ₁₆ is H; C _1-6 alkyl, C _3-7 cycloalkyl, or C _{4 -10} alkylcycloalkyl (respectively, halo, C _1-3 alkoxy, sometimes C _1-3 haloalkoxy, substituted with C _1-3 alkyl or C _1-3 haloalkyl); is aryl or Het ;
(c) R ₈ and R ₉ are each independently C _1-3 alkyl, or C _1-3 alkoxy, or C _1-3 haloalkoxy, which may be substituted with H, or halo;
(d) Q is a saturated or unsaturated C _3-9 chain that may contain 1 to 3 heteroatoms independently selected from O, S (O) _m ; (where m is 0, 1 , Or 2), or NR ₁₇ where R ₁₇ is H; C _1-6 alkyl or C _1-6 cycloalkyl (each halo, C _1-6 alkoxy, cyano, or C _1-6 halo) May be substituted with alkoxy); —C (O) —R ₁₈ , C (O) —OR ₁₉ , C (O) —NR ₂₀ R ₂₁ , or —SO ₂ R ₂₂ ; R ₁₈ , R ₂₀ and R ₂₁ are each independently H; C _1-6 alkyl, or C _1-6 cycloalkyl (each substituted with halo, C _1-6 alkoxy, cyano, or C _1-6 haloalkoxy R ₁₉ is C _1-6 alkyl or C _1-6 cycloalkyl (each substituted with halo, C _1-6 alkoxy, cyano, or C _1-6 haloalkoxy) May be);
R ₂₂ is aryl, C _1-6 alkyl, or C _1-6 cycloalkyl, each optionally substituted with halo, C _1-6 alkoxy, cyano, or C _1-6 haloalkoxy. ;
(e) W is OH, —NH—SO _n —R ₂₃ , or NH—SO _n —R ₂₄ ; (where n is 1 or 2), R ₂₃ is C _1-8 alkyl, C _4-10 alkyl cycloalkyl, unsubstituted C _3-7 cycloalkyl, or halo, C _1-3 alkoxy, cyano, amine, mono or di-C _1-6 alkyl amine, mono or di-C _1-6 alkyl Amido, or C _7-9 alkylaryl optionally substituted with carboxylate or cyclopropyl or cyclobutyl optionally substituted with C _1-4 alkyl; R ₂₄ is C _6-10 aryl or Het It is. ]. R ₁ is bonded to the C ₃ position; H; C _1-6 alkyl; C _3-7 cycloalkyl; C _1-6 alkoxy; C _3-7 cycloalkoxy; halo-C _1-6 alkoxy; halo-C _{1 -6} alkyl; cyano; halo; C _1-6 alkanoyl; mono- or di- (C _1-6 ) alkylamine; mono- or di-C _1-6 alkylamide; carboxyl; may be substituted with Het A compound according to claim 1 selected from _6-10 aryl; C _7-14 alkylaryl; C _6-10 aryloxy or Het. R ₂ , R ₃ , and R ₄ are bonded to the C ₄ , C ₅ , and C ₆ positions, respectively, and are independently H; C _1-6 alkyl; C _3-7 cycloalkyl; C _1-6 alkoxy; C _3-7 cycloalkoxy; halo-C _1-6 alkoxy; halo-C _1-6 alkyl; cyano; halo; hydroxy; C _1-6 alkanoyl; mono- or di- (C _1-6 ) alkylamine; C ₆ which may be substituted with Het; carboxyl; di - - (C _3-7) cycloalkyl amine; mono- or di -C _1-6 alkyl amide (C _3-7) cycloalkyl amide mono- or di _2. The compound of claim 1, selected from: _-10 aryl; C _7-14 alkyl aryl; C _6-10 aryloxy; or Het. R ₅ and R ₆ are bonded to the C ₇ and C ₈ positions, respectively, and are independently H; C _1-3 alkyl; C _3-4 cycloalkyl; C _1-3 alkoxy; C _3-4 cycloalkoxy; -C _1-3 alkoxy; halo-C _1-3 alkyl; cyano; halo; hydroxy; C _1-3 alkanoyl; mono- or di- (C _1-3 ) alkylamine; mono- or di- (C _3-4 ) cycloalkyl amine; mono- or di -C _1-3 alkyl amide; mono- or di - (C _3-4) cycloalkyl amide; or a compound according to claim 1 selected from carboxyl. A saturated or unsaturated C _3-9 chain in which Q may contain 1 to 3 heteroatoms independently selected from O, S (O) _m ; (where m is 0, 1, or 2; there), or a NR _17, R ₁₇ is H; C _1-6 alkyl, C _{1-6 cycloalkyl, -C (O) -R 18,} C (O) -OR 19, C (O) -NR _2. The compound according to claim 1, which is ₂₀ R ₂₁ or —SO ₂ R ₂₂ . R ₁₈ , R ₂₀ , and R ₂₁ are each independently H; C _1-6 alkyl or C _1-6 cycloalkyl; R ₁₉ is C _1-6 alkyl or C _1-6 cycloalkyl 6. The compound of claim 5, wherein R ₂₂ is aryl, C _1-6 alkyl, or C _1-6 cycloalkyl, each of which may be substituted with halo. W is OH, -NH-SO _n -R ₂₃ , or NH-SO _n -R ₂₄ , n is 1 or 2, and R ₂₃ is unsubstituted C _3-7 cycloalkyl, or C _7-9 alkyl it is cyclopropyl or cyclobutyl is substituted with aryl or C _1-4 alkyl the compound according to claim 1, wherein R ₂₄ is C _6-10 aryl or Het. Formula II:

Or a pharmaceutically acceptable enantiomer, diastereomer, salt, solvate, or prodrug thereof [wherein
(a) R ₁ is H; C _1-6 alkyl; C _3-7 cycloalkyl; C _1-6 alkoxy; C _3-7 cycloalkoxy; halo-C _1-6 alkoxy; halo-C _1-6 alkyl cyano, halo, C _1-6 alkanoyl; mono- or di - (C _1-6) alkyl amine; C _6-10 that may have been substituted with Het; mono- or di -C _1-6 alkyl amide; carboxyl Aryl; C _7-14 alkylaryl; C _6-10 aryloxy or Het; the R ₁ may be linked to the isoquinoline group by a C _1-6 alkyl linking group;
R ₂ , R ₃ , and R ₄ are each independently H; C _1-6 alkyl; C _3-7 cycloalkyl; C _1-6 alkoxy; C _3-7 cycloalkoxy; halo-C _1-6 alkoxy Halo-C _1-6 alkyl; cyano; halo; hydroxyl; C _1-6 alkanoyl; mono- or di- (C _1-6 ) alkylamine; mono- or di- (C _3-7 ) cycloalkylamine; Di-C _1-6 alkylamide; mono- or di- (C _3-7 ) cycloalkylamide; carboxyl; C _6-10 aryl optionally substituted with Het; C _7-14 alkylaryl; C _{6- 10} aryloxy; or Het; the R _{2 to} R ₄ may be linked to the isoquinoline group by a C _1-3 alkyl linking group; R ₅ and R ₆ are each independently H; C _{1 -3} alkyl; C _3-4 cycloalkyl; C _1-3 alkoxy; C _3-4 cycloalkoxy; halo-C _1-3 alkoxy; halo-C _1-3 a Cyano; halo; hydroxyl; C _1-3 alkanoyl; mono- or di- (C _1-3 ) alkylamine; mono- or di- (C _3-4 ) cycloalkylamine; mono- or di-C _1-3 alkyl Amide; mono- or di- (C _3-4 ) cycloalkylamide; or carboxyl;
(b) R ₇ is NH ₂ or —NR ₁₀ R ₁₁ ; R ₁₀ is C _1-6 alkyl, C _1-6 haloalkyl, C (O) —NR ₁₂ R ₁₃ , C (O) —OR ₁₄ , or is _{-C (O) -R 16; R} 11 is H, is C _1-6 alkyl or C _1-6 haloalkyl; provided that when either R ₁₂ or R ₁₃ is H R ₁₁ R ₁₂ and R ₁₃ are each independently H; C _1-6 alkyl, C _3-7 cycloalkyl, or C _4-10 alkyl cycloalkyl (each halo, C _1-3 alkoxy, May be substituted with C _1-3 haloalkoxy, C _1-3 alkyl, or C _1-3 haloalkyl); R ₁₂ and R ₁₃ together with the nitrogen to which they are attached 4-14 membered heterocycles may be formed; R ₁₄ and R ₁₅ are each independently C _1-6 alkyl, C _3-7 cycloalkyl, or C _4-10 alkyl cycloalkyl (each halo, C _1-3 alkoxy, C _1-3 haloalkoxy, C _1-3 Alkyl or it is there) substituted with C _1-3 haloalkyl,; R ₁₆ is, H; C _1-6 alkyl, C _3-7 cycloalkyl or C _4-10 alkylcycloalkyl (each, May be substituted with halo, C _1-3 alkoxy, C _1-3 haloalkoxy, C _1-3 alkyl, or C _1-3 haloalkyl); aryl or Het;
(c) R ₈ and R ₉ are each independently C _1-3 alkyl, or C _1-3 alkoxy, or C _1-3 haloalkoxy, which may be substituted with H, or halo;
(d) Q is a saturated or unsaturated C _3-9 chain that may contain 1 to 3 heteroatoms independently selected from O, S (O) _m ; (where m is 0 , 1 or 2), or NR ₁₇ where R ₁₇ is H; C _1-6 alkyl, C _1-6 cycloalkyl, —C (O) —R ₁₈ , C (O) —OR ₁₉ , C (O) —NR ₂₀ R ₂₁ , or —SO ₂ R ₂₂ ; R ₁₈ , R ₂₀ , and R ₂₁ are each independently H; C _1-6 alkyl or C _1-6 R ₁₉ is C _1-6 alkyl or C _1-6 cycloalkyl; R ₂₂ is aryl, C _1-6 alkyl, or C _1-6 cycloalkyl, each substituted with halo May be);
(e) W is OH, —NH—SO _n —R ₂₃ , or NH—SO _n —R ₂₄ , where n is 1 or 2; R ₂₃ is unsubstituted C _3-7 cyclo Alkyl, or cyclopropyl or cyclobutyl optionally substituted with C _7-9 alkylaryl or C _1-4 alkyl; R ₂₄ is C _6-10 aryl or Het. ]. R ₁ is substituted with H; C _1-3 alkoxy; mono- or di- (C _1-6 ) alkylamine; 5- or 6-membered monocyclic heterocycle; or 5- or 6-membered monocyclic heterocycle _9. The compound of claim 8, which is _optionally C _6-10 aryl. R ₂ , R ₃ , R ₄ , and R ₅ are each independently H; C _1-6 alkoxy; halo-C _1-6 alkoxy; hydroxyl; or mono- or di- (C _1-6 ) alkylamine 9. The compound according to claim 8. R ₇ is NH ₂ or —NHR ₁₀ ; R ₁₀ is C (O) —NR ₁₂ R ₁₃ or C (O) —OR ₁₄ ; R ₁₂ and R ₁₃ may be substituted with halo C _1-6 is alkyl; R ₁₄ is be substituted with halo C _1-6 alkyl or C _3-7 compound of claim 8, wherein cycloalkyl. Q is a C _5-7 membered chain having one double bond that may contain one heteroatom independently selected from O, S (O) _m (where m is 0, 1 Or is 2), or NR ₁₇ (wherein R ₁₇ is H; C _1-6 alkyl or C _1-6 cycloalkyl). 9. The compound of claim 8, wherein Q has the following structure:

Wherein P is a saturated C ₃ chain that may contain one hetero atom independently selected from O, S (O) _m, where m is 0, 1, or 2. Or NR ₁₇ . ]. W is —NH—SO _n —R ₂₃ (where n is 1 or 2) and R ₂₃ is unsubstituted C _3-7 cycloalkyl, or C _7-9 alkylaryl or C _1-4 alkyl 9. The compound according to claim 8, which is cyclopropyl or cyclobutyl which may be substituted with. 9. The compound according to claim 8, wherein W is NH—SO _n —R ₂₄ , n is 1 or 2, and R ₂₄ is Het. The Het

16. The compound according to claim 15, selected from the group consisting of: Formula III:

Or a pharmaceutically acceptable enantiomer, diastereomer, salt, solvate, or prodrug thereof [wherein
(a) R ₁ is H; C _1-3 alkoxy; di- (C _1-6 ) alkylamine; 5- or 6-membered monocyclic heterocycle; or 5- or 6-membered monocyclic heterocycle is is C _6-10 aryl that may occur _{are; R 2, R 3, R} 4, and R ₅ are each independently H; C _1-3 alkoxy; halo; or di - (C _1-6 Alkylamine;
(b) R ₇ is —NHR ₁₀ ; where R ₁₀ is C (O) —NHR ₁₃ or C (O) —OR ₁₄ ; R ₁₃ and R ₁₄ are C _1-6 alkyl;
(c) Q is a C _5-7 membered chain having one double bond that may contain one heteroatom independently selected from O and S (O) _m (where m is Is 0, 1, or 2), or NR ₁₇ where R ₁₇ is H; C _1-6 alkyl or C _1-6 cycloalkyl;
(d) R ₂₃ is unsubstituted C _3-7 cycloalkyl, or cyclopropyl or cyclobutyl which may be substituted with C _7-9 alkylaryl or C _1-4 alkyl. ]. 18. The compound of claim 17, wherein R ₁ is selected from the group consisting of pyridine, morpholine, piperazine, oxazole, isoxazole, thiazole, imidazole, pyrrole, and pyrazole. R ₁ is 1 or selected from the group consisting of C _1-3 alkoxy, halo, carboxyl, di- (C _1-3 ) alkylamine, C _1-3 haloalkyl, trifluoromethyl, trifluoromethoxy, and hydroxy 18. A compound according to claim 17, which is phenyl which may be substituted with the above members. 18. The compound according to claim 17, wherein R _{1 is} di- (C _1-3 ) alkylamine. 18. The compound of claim 17, wherein R ₁ is piperazine substituted with one or more members selected from the group consisting of C _1-3 alkyl, C _5-7 cycloalkyl, or pyridine. The compound of claim 17 wherein R ₂ is chloro or fluoro. The compound according to claim 17, wherein R _{2 is} di- (C _1-3 ) alkylamine or methoxy. Q is

18. The compound according to claim 17, having a structure selected from: Q is

18. The compound according to claim 17, having a structure selected from:

A compound selected from the group consisting of:   A composition comprising the compound of claim 1 and a pharmaceutically acceptable carrier.   28. The composition of claim 27, further comprising a compound having anti-HCV activity.   29. The composition according to claim 28, wherein the compound having anti-HCV activity is interferon.   30. The composition of claim 29, wherein the interferon is selected from the group consisting of interferon alpha 2B, polyethylene glycol pegylated interferon alpha, consensus interferon, interferon alpha 2A, and lymphoblastiod interferon tau.   Compounds with anti-HCV activity enhance interleukin-2, interleukin-6, interleukin-12, type 1 helper T-cell expression, interfering RNA, antisense RNA, imiqimod, ribavirin, inosine 5'-monophosphate dehydrogenase 29. The composition of claim 28, selected from the group consisting of an inhibitor, amantadine, and rimantadine.   29. The composition of claim 28, further comprising interferon and ribavirin.   29. The composition according to claim 28, wherein the compound having anti-HCV activity is a small molecule compound.   Compounds with anti-HCV activity can be used to treat HCV infection, HCV metalloprotease, HCV serine protease, HCV polymerase, HCV helicase, HCV NS4B protein, HCV entry, HCV assembly, HCV release, HCV NS5A protein, IMPDH, 29. The composition of claim 28, which is effective to inhibit the function of a target selected from the group consisting of and nucleoside analogs.   A method for inhibiting the function of HCV serine protease, which comprises contacting the compound of claim 1 with HCV serine protease.   A method of treating HCV infection in a patient comprising administering to the patient a therapeutically effective amount of a compound according to claim 1, or a pharmaceutically acceptable enantiomer, diastereomer, solvate, prodrug, or salt thereof. .   38. The method of claim 36, wherein the compound is effective to inhibit the function of HCV serine protease.   38. The method of claim 36, further comprising administering another compound having anti-HCV activity before, after, or simultaneously with the compound of claim 1.   39. The method of claim 38, wherein the other compound having anti-HCV activity is interferon.   40. The method of claim 39, wherein the interferon is selected from the group consisting of interferon alpha 2B, polyethylene glycol-added interferon alpha, consensus interferon, interferon alpha 2A, lymphoblastoid interferon tau.   Other compounds with anti-HCV activity include interleukin 2, interleukin 6, interleukin 12, compounds that enhance the expression of type 1 helper T cell response, interfering RNA, antisense RNA, imiqimod, ribavirin, inosine 5'- 40. The method of claim 38, selected from the group consisting of a monophosphate dehydrogenase inhibitor, amantadine, and rimantadine.   39. The method of claim 38, wherein the compound having anti-HCV activity is a small molecule.   Compounds with anti-HCV activity can be used to treat HCV infection, HCV metalloprotease, HCV serine protease, HCV polymerase, HCV helicase, HCV NS4B protein, HCV entry, HCV assembly, HCV release, HCV NS5A protein, IMPDH, 43. The method of claim 42, wherein the method is effective to inhibit the function of a target selected from the group consisting of and a nucleoside analog.   39. The method of claim 38, wherein the other compound having anti-HCV activity is effective to inhibit the function of a target in the HCV life cycle other than HCV serine protease.   Use of a compound according to claim 1 for the manufacture of a medicament for treating HCV infection in a patient.   28. Use of the composition according to claim 27 for the manufacture of a medicament for treating HCV infection in a patient.   A method of resolving a mixture of alkyl ester enantiomers, the method comprising contacting the mixture with an enzyme effective to selectively promote hydrolysis of one of the enantiomers in the presence of a buffering agent. A method characterized by being performed in The alkyl ester has the following formula:

[Wherein R ₂₅ is an amino protecting group;
R ₂₆ is selected from the group consisting of C _1-10 alkyl, C _6-14 aryl, C _7-16 alkylaryl, C _3-7 cycloalkyl, or C _3-10 alkylcycloalkyl. ]
48. The method of claim 47, comprising:   48. The method of claim 47, wherein the buffer is selected from the group consisting of phosphate, borate, and carbonate.   48. The method of claim 47, wherein the enzyme is a protease.   48. The method of claim 47, wherein the enzyme is selected from the group consisting of Bacillus globigii, Bacillus licheniformis, Bacillus halodurans, Bacillus clausii, Aspergillus oryzase, and mixtures thereof.   The enzyme is Alcalase (registered trademark) (subtilisin protease), Savinase (registered trademark) (subtilisin protease), Esperase (registered trademark) (subtilisin protease), Flavorzyme (registered trademark) (fungal protease), 52. The method of claim 51, wherein the method is a mixture thereof.   48. The method of claim 47, wherein the contacting is performed at a pH of about 7.0-11.   48. The method of claim 47, wherein the contacting is performed at a temperature of about 30-60 ° C.   48. The method of claim 47, wherein the contacting is performed for a time of less than about 7 days.   56. The method of claim 55, wherein the contacting is performed for a period of about 2 hours to 3 days.